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| Hematology Urology Blood Bank Microbiology Cytology/Histology Miscellaneous |
METHANOL FIXATIVE For use as a fixative prior to staining bloody specimen material or blood culture supematant fluid. Methanol preserves the morphology of red blood cells as well as bacteria. RECOMMENDED PROCEDURE: Place material to be stained on a clean glass slide. Flood slide with methanol fixative for 1 minute. Drain without rinsing and allow slide to air dry prior to staining.
CONTENTS:
Methanol ETHANOL FIXATIVE, 80% Fixation with ethanol is useful for examining bloody specimen material or blood culture supernatant fluid. Ethanol preserves the morphology of red blood cells as well as bacteria. RECOMMENDED PROCEDURE: The fixation of material on a glass slide prior to staining may be accomplished by dipping or flooding for up to five minutes.
CONTENTS:
Ethanol WRIGHT, WRIGHT GIEMSA AND MAY-GRUNWALD GIEMSA STAINS NOTE: Due to the variations in the oxidation of a methylene blue stain, our Wright, Wright Giemsa and May-Grunwald Giemsa stain products are offered with a matched buffer solution of specific pH in order to obtain consistent and uniform staining characteristics from lot to lot. As a result, we recommend our matched buffer solution be used for obtaining optimum results. However, stain and buffer are available separately. H-PACK* STAIN PACKS H-PACK* II STAIN PACK These stain packs are designed to eliminate problems with precipitate, artifacts, drying of slides, and overstaining which causes abnormal staining characteristics of all cellular elements, i.e., false toxic granulation. One adjustment should be all that is necessary to obtain desired color and intensity. Buffer solution compliments individual stain lots to allow consistent staining characteristics. Normal and abnormal WBC’s and platelets are vividly demonstrated. Bacteria within leukocytes can be clearly observed. RBC staining gives excellent results of normal cellular elements including all abnormalities and inclusion bodies. Parasites, such as malaria, are well-defined. Bone marrow preps can be stained routinely. Freezing or high temperature will not cause change in staining ability. Stain pack is designed to be used at room temperature. SHAKE WELL BEFORE USE. RECOMMENDED USES: H-PACK* & H-PACK*II may be used on all specimens requiring a WBC differential, particularly spinal fluid and nasal smears, and any procedure requiring a Wright counterstain. Other uses include LE preps, parasitic elements such as malaria, thicker buffy coat smears, mature and immature elements of routine bone marrow smears, inclusion bodies such as basophilic stippling, etc. All stain clearly and distinctly. RECOMMENDED PROCEDURE: Prior to inserting stain pack, all cannulas, tubing and platen surfaces should be cleaned with methanol. For optimum results, new tubing should be soaked in methanol. In order to prevent staining inconsistencies or artifacts due to quality of slides or humid conditions affecting the smear, we recommend fixing in methanol. (After fixing, it is not necessary to dry slide prior to insertion into machine.) Crenation of red blood cells can be caused by mechanical changes in preparation of slide prior to staining. H-PACK*: Place stain pack into your instrument with the cut out holes facing forward. Insert cannulas through plastic bottles until flush. NOTE: If cannulas are dull and do not puncture bottles smoothly, use an 18 - 20 gauge needle to puncture bottles prior to inserting cannulas. H-PACK* II: The cannulas are inserted into the appropriate containers of solution. The stain container has two cannulas, one inserted at each end of the container. The buffer and rinse containers have one cannula each. RECOMMENDATION FOR MACHINE SETTINGS: Due to the inconsistency in the pump motors of the Ames HEMA-TEK** Slide Stainers and the constant wear of the pump tubing, we recommend that a daily setting be made in order to obtain consistent results from day to day. H-PACK* WRIGHT, WRIGHT GIEMSA AND MAY-GRUNWALD GIEMSA STAINS: Ames Hema-Tek** and Ames Hema-Tek ** 1000: All pumps should initially be set on maximum by rotating the pumps completely to the right. For a more acid (red) stain, keep buffer and rinse pumps on full and reduce stain pumps by rotating setting to the left. For a more alkaline (blue) stain, keep stain and rinse pumps on full and reduce buffer by rotating setting to the left. Ames Hema-Tek** 2000: All pumps should initially be set to +2. For a more acid (red) stain, adjust only stain pump by rotating setting to 0 or -2. For a more alkaline (blue) stain, adjust only buffer pump by rotating setting to 0 or -2. Due to the unstable characteristics of formulating a stain pack that contains both a Wright, Giemsa or May-Grunwald stains, the presence of some residue on the platen will be seen after use. It is suggested that the platen surface be cleaned at frequent intervals to prevent build-up of stain residue on the platen. H-PACK* II WRIGHT GIEMSA STAIN: Since the H-PACK* II stain requires a shorter staining time, both quantities of solution and time may be decreased. Setting the rinse down to the minimum, stain volume and stain intensity control may be decreased yielding excellent staining quality. NOTE: RINSE SHOULD BE SET ON FULL AT ALL TIMES TO AID IN CLEANING AND DRYING OF SLIDES. TROUBLESHOOTING COMMON STAINING PROBLEMS: Ames HEMA-TEK** Slide Stainers: Change all tubing monthly and flush stain line with methanol. Use 25 gauge needle and syringe and place in stain opening on platen. Remove stain pump tubing and check to determine if methanol is flushing out from rear connector. Clean platen and surrounding wells. Remove all debris and be careful not to bend finger switches. Make sure finger switches are clean and floating freely. Run blank slide through stainer and check pump times. Stain pump should be activated when center of slide is over opening on platen and pump should run for about 8 seconds. Be sure the slide is completely covered with the stain. Buffer pump should be activated when edge of slide is over opening on platen and pump should run for about 45 seconds. Rinse pump should be activated when edge of slide is over opening on platen. This should run for 40 - 45 seconds and pump should remain on full. Adjust finger switches to correct. * Trademark of
ENG Scientific, Inc. PRECAUTIONS: The solvent for the stain is methanol, which is flammable. Store at room temperature. IN THE EVENT OF COLD WEATHER, allow solution to return to room temperature and SHAKE WELL BEFORE USE.
CONTENTS: Wright Stain, Giemsa Stain, May-Grunwald Stain, Methanol, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable).
RECOMMENDED PROCEDURE FOR MIDAS, HEMASTAINER, AND CARL ZEISS HMS SLIDE STAINERS: Fill Station 1 with Traxstain Fixative. Fill Station 2 with Traxstain (Wright, Wright Giemsa or May- Grunwald Giemsa stain). Fill station 3 by mixing 80 ml of Traxstain with 420 ml of Buffer. Station 4 is a rinse station as provided for the instrument. Fill station 5 with Traxstain Buffer. RECOMMENDED PROCEDURE FOR MIDAS II, AND SAKURA RSG-61 SLIDE STAINERS: Fill Station 1 with Traxstain Fixative. Fill Station 2 with Traxstain (Wright, Wright Giemsa or May-Grunwald Giemsa Stain). Fill station 3 by mixing 50 ml of Traxstain with 250 ml of Buffer. Station 4 is a rinse station as provided for the instrument. Fill station 5 with Traxstain Buffer.
--------------SUGGESTED
TIMES-------------
NOTE: These are median times and may be adjusted for desired intensity, keeping a ratio of 1:2 for Station 2 and 3. The rinse water must be within the range of pH 6.5 to 7.2. In order to obtain proper staining results, we suggest using the minimum time setting for this station, i.e. 5 seconds. Stain color is adjusted by increasing or decreasing time in Station Number 5 (Traxstain Buffer); less time, more alkaline (blue), more time, more acidic (red). IN THE EVENT OF COLD WEATHER, allow solution to return to room temperature and SHAKE WELL BEFORE USE.
An alcoholic quick stain technique (fixation not necessary) requiring less than one minute yielding excellent staining results without precipitate. RECOMMENDED PROCEDURE: Pour solutions into three separate coplin jars. Place slide in Sol. I for 10 seconds. DO NOT AGITATE. Place in Sol. II for 10 seconds. DO NOT AGITATE. Place in Sol. III for 10 seconds. DO NOT AGITATE. For adequate rinsing, dip slide in and out of Sol. III approximately 3 - 4 times. Allow to dry. NOTE: Less time in Sol. III provides a more alkaline (blue) stain; increase time for a more acidic (red) stain. IN THE EVENT OF COLD WEATHER, allow solutions to return to room temperature and SHAKE WELL BEFORE USE. * Trademark of Eng Scientific, Inc.
CONTENTS: Wright Stain, Giemsa Stain, Methanol, Buffers, Stabilizers, Eosin These
solutions are made from certified dyes (when applicable). STAT-QUICK
STAIN No fixation required with this stain. Utilized in both stat and routine laboratories. Only two solutions are necessary to obtain excellent staining characteristics. Matching buffer solution allows uniform staining of all cellular elements without precipitate or artifactual changes. RECOMMENDED PROCEDURE: Place slide in stain for 20 seconds. DO NOT AGITATE. Place slide in buffer (water, if used, see NOTE) for 20 seconds depending upon color desired. DO NOT AGITATE. For a more alkaline (blue) stain, decrease time in buffer solution. For a more acid (red) stain, increase time in buffer solution. To facilitate adequate cleansing of slide, shake 3 - 4 times in buffer solution. Allow to dry. NOTE: For more intense staining, increase time in stain and buffer solution. We have noted that in some areas the tap water as well as distilled water varies in pH.This will result in improper staining. We recommend that our buffer solution be used to maintain consistency in staining quality. If water is used, check pH of the water to be in the range of 6.7 to under 6.8. STAIN CONTAINER SHOULD ALWAYS BE SHAKEN BEFORE USE, and the stain placed in the staining container should be mixed periodically. IN THE EVENT OF COLD WEATHER, allow solution to return to room temperature and SHAKE WELL BEFORE USE.
CONTENTS: Wright Stain, Giemsa Stain, Methanol, Eosin, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable). MANUAL METHOD
STAIN This solution, using a flooding technique, will stain all cellular elements without precipitate or artifactural changes. Staining characteristics are similar to H-PACK*. Buffer is applied immediately after stain, reducing overall staining time. Convenient flip-top cap for 250 ml and 950 ml sizes. IN THE EVENT OF COLD WEATHER, allow solution to return to room temperature and SHAKE WELL BEFORE USE. RECOMMENDED PROCEDURE: We recommend slides be fixed in methanol. Flood slide with stain. Immediately, add equal amount of buffer to fully cover and mix with stain. (Blow gently on slide to adequately mix both buffer and stain.) Stain 5 - 7 minutes depending on color intensity desired. If you desire a more alkaline (blue) stain, decrease staining time. If you prefer a more acid (red) stain, increase staining time. Rinse slide with distilled water and allow to dry before reading.
CONTENTS: Wright Stain, Giemsa Stain, Methanol, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable). METHANOL FIXATIVE For use as a fixative prior to staining bloody specimen material or blood culture supematant fluid. Methanol preserves the morphology of red blood cells as well as bacteria. RECOMMENDED PROCEDURE: Place material to be stained on a clean glass slide. Flood slide with methanol fixative for 1 minute. Drain without rinsing and allow slide to air dry prior to staining.
CONTENTS: Methanol FOR IN VITRO DIAGNOSTIC USE ONLY. DIP STAIN KIT Solution I - Fixative RECOMMENDED PROCEDURE: Fix slide in Sol. I for 30 seconds. Place in Sol. II for 3 minutes followed by Sol. III for 1 minute. NOTE: The color can be varied by either increasing or decreasing time in Sol. III (Matching Buffer). For a more basic stain (blue), decrease time in Sol. III; for a more acid stain (red), increase time. Dip slide in Sol. IV for 3 - 5 seconds. NOTE: Due to evaporation and absorption of moisture, it may be necessary to increase time in Sol. I (Fixative) to 1 minute, Sol. II (Stain) to 3.5 minutes, Sol. IV (Alcohol Wash) may be increased to 30 seconds. Solutions should be changed after a 24 hour period with cleaning of all reagent containers. Solutions should be replenished during staining period. IN THE EVENT OF COLD WEATHER, allow solutions to return to room temperature and SHAKE WELL BEFORE USE.
CONTENTS: Wright Stain, Giemsa Stain (as indicated), Methanol, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable). GIEMSA STAIN RECOMMENDED PROCEDURE: For use as a counterstain, dilute stock Giemsa with an equal volume of buffer or distilled water. We suggest a 20 minute staining time. If used as a straight Giemsa stain, increase ratio of stain to buffer for darker staining. NOTE: We recommend a buffer solution within a 6.8 pH range or distilled water with a comparable pH range.
CONTENTS: Giemsa Stain, Methanol This solution
is made from certified dyes. WOLBACH'S GIEMSA STAIN A stain used to stain bone marrow preparations and to aid in the identification of bacteria, rickettsia, and collagen. RECOMMENDED PROCEDURE: Deparaffinize and hydrate to distilled water. Remove mercuric chloride crystals with iodine and clear with sodium thiosulfate. Wash in running water for 15 minutes. Rinse in distilled water. Working Wolbach's Giemsa solution* overnight. Differentiate in working rosin alcohol solution until sections assume a purplish pink color. Check microscopically. Dehydrate in absolute alcohol then clear in xylene, two changes each. Mount with Permount or Histoclad. *Working Solution: Distilled water - 50 ml, Methanol - 1.5 ml, Wolbach's Giemsa - 1.25 ml.
CONTENTS: Giemsa Stain, Methanol, Glycerin This solution
is made from certified dyes.
NATT-HERRICK'S
STAIN A solution that stains all white blood cells dark violet distinguishing them from red blood cells. RECOMMENDED PROCEDURE: Draw venous or capillary blood to the 1.0 mark in a white cell pipette. Draw eosinophil solution to the 11.0 mark and mix gently for 30 seconds. NOTE: Prolonged, vigorous shaking will tend to cause rupturing of eosinophils. Allow to stand between 5 - 15 minutes. Shake again, charge chamber and allow cells to settle before counting. 1. Prepare a 1:200 dilution of blood with Natt-Herrick's stain (add 20 ul blood to 4 ml of Natt-Herrick's stain). 2. Mix well, and leave at room temperature for 5 minutes: then fill both sides of a hemocytometer with the stained blood. 3. After 5 minutes, perform a white blood cell count, using 10X objective. That is, count all white blood cells in the 4 large corner squares on both sides of the hemocytometer chamber (the counts within each square should be within 10 percent of each other). Add all 8 counts together and use this total count to calculate.
Total # WBC's X 2000 = # WBC's/ul blood All white blood cells will stain dark violet.
CONTENTS: Sodium chloride, Sodium sulfate, Sodium phosphate, Potassium phosphate, Formalin, Methyl Violet. This solution is made
from certified dyes. EOSINOPHIL DILUTING FLUID AND STAIN Eosinophil counts are useful in determining an increase (eosinophilia) or decrease (eosinopenia) in eosinophilic granulocytes. This technique, used to perform an absolute count, requires a special diluent and/or stain solution. This diluting fluid will lyse the erythrocytes and only eosinophils will stain a bright orange-red. RECOMMENDED PROCEDURE: Draw venous or capillary blood to the 1.0 mark in a white cell pipette. Draw eosinophil solution to the 11.0 mark and mix gently for 30 seconds. NOTE: Prolonged, vigorous shaking will tend to cause rupturing of eosinophils. Allow to stand between 5 - 15 minutes. Shake again, charge chamber and allow cells to settle before counting. Cells Counted x Dilution (10) x
Depth (10) = Total Cells / cmm
CONTENTS: Phloxine, Propylene Glycol, Sodium Carbonate This solution is made
from certified dyes.
EOSINOPHIL
DILUTING FLUID AND STAIN A diluting fluid that renders the red blood cells invisible (hemolyzes) and lyses all WBC's except eosinophils. RECOMMENDED PROCEDURE: Draw venous or capillary blood to the 1.0 mark in a white cell pipette. Draw eosinophil solution to the 11.0 mark and mix gently for 30 seconds. NOTE: Prolonged, vigorous shaking will tend to cause rupturing of eosinophils. Allow to stand between 5 - 15 minutes. Shake again, charge chamber and allow cells to settle before counting. 1. Draw venous or capillary blood to the 1.0 mark in a white cell pipette. 2. Draw eosinophil solution to the 11.0 mark and mix gently for 30 seconds. NOTE: Prolonged, vigorous shaking will tend to cause rupturing of eosinophils. 3. Allow to stand between 5 - 15 minutes. 4. Shake again, charge chamber and allow cells to settle before counting. Cells Counted x Dilution (10) x
Depth (10) = Total Cells / cmm
CONTENTS: Phloxine, Propylene Glycol, Sodium Carbonate Phloxine, Propylene Glycol, Sodium Carbonate. This solution is made
from certified dyes. URINARY
SEDIMENT STAIN A quick staining procedure by which a drop of stain solution is directly added to urine sediment. Neutrophilic leukocytes stain violet with red-purple nuclei. "Glitter" cells stain light blue to almost colorless. Squamous vaginal epithelial cells stain pale purple; nucleus stains dark purple. Bladder epithelial cells are either colorless or a pale blue. Hyaline casts stain a delicate pink to rose shade. Granulation stains red violet, or blue in granular casts. WBC, REC, epithelial, and cellular casts are easily recognized by the intense staining characteristics of cellular inclusions. Fatty cells have a bright honeycomb-like structure in a slightly stained matrix. Bacteria stain pink when living and active; dark purple when dead. Yeast cells may stain dark purple or they may not take the stain at all. Trichomonas parasites are either colorless or pale blue. RBC’s stain faintly.
CONTENTS: Crystal Violet, Ammonium Oxalate, Ethanol, Safranin O This solution
is made from certified dyes. PRUSSIAN
BLUE STAIN KIT Solution I - Potassium
Ferrocyanide (2%) RECOMMENDED PROCEDURE: Working solution: Just prior to use, mix 5 ml of Sol. I with 5 ml of Sol. II. Hemosiderin in urine appears as yellow-brown granules seen either free or within epithelial cells and occasionally in casts. Examine several drops of a centrifuged urine specimen for coarse brown granules. If coarse brown granules are present, resuspend the remainder of the sediment in the working solution and allow to remain for 10 minutes. Centrifuge specimen, discard supematent and examine microscopically. NOTE: The use of chemically clean glassware and plastic-tipped or paraffin-coated forceps is suggested. RESULTS: Coarse granules of hemosiderin stain blue.
CONTENTS: Potassium Ferrocyanide, HCl For in vitro diagnostic use only. MALARIAL
WRIGHT GIEMSA STAIN KIT Solution I - Fixative This stain kit (a modified Wright Giemsa formulation) utilizes an alcohol fixative and stain in combination with a buffer solution (pH range - 6.8 to 7.2). All cellular elements are stained. The use of a thin smear allows for easy visualization of inclusions, extracellular forms and the size of red cells. Their characteristic morphologies are used for differentiation. RECOMMENDED PROCEDURE: Make a thin blood smear. Fix slide in Sol. I from 10 - 30 seconds. Flood slide with Sol. II. Immediately overlay with equal volume of Sol. III. Stain 3 - 6 minutes depending upon color intensity desired. Rinse and allow slide to dry in a vertical position.
CONTENTS: Wright Stain, Giemsa Stain, Methanol, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable). MALARIAL
QUICK STAIN KIT Solution I - Buffered Stain A combination of water-based buffered stains (pH range = 6.8 to 7.2) whereby the RBC’s are hemolyzed allowing the malarial parasites to be stained. Thick films provide a greater volume of blood and therefore, larger numbers of organisms for examination. However, characteristic definitive morphologic criteria may be more difficult to see. The cytoplasm of white blood cells will stain with eosinophils predominant. RECOMMENDED PROCEDURE: Allow blood film to air dry completely. DO NOT FIX IN ALCOHOL OR HEAT. Place enough of Sol. I and Sol. II into two separate coplin jars. Place unfixed blood smear in Sol. I for 1 - 3 seconds. Remove and rinse under tap water until all stain is removed. Place in Sol. II for 2 seconds. Rinse under tap water 2 - 3 seconds and allow to dry.
CONTENTS: Methylene Blue, Giemsa Stain, Eosin Y, Buffers, Stabilizers These
solutions are made from certified dyes (when applicable). MAY- GRUNWALD
STAIN KIT Solution I - May-Grunwald Stain RECOMMENDED PROCEDURE: Fix smears for 3 minutes in methanol. Cover slide with Sol. I for 3 minutes. Add an equal amount of distilled water and allow to stand 1 minute. Drain without rinsing. Cover for 12 minutes with dilute Giemsa stain (15 drops of Sol. II + 10 drops of distilled water.) Differentiate in distilled water by agitating for about 5 seconds and checking under microscope. Blot dry and mount. RESULTS: Similar to a Wright Giemsa stain.
CONTENTS: May-Grunwald Stain, Giemsa Stain, Methanol These
solutions are made from certified dyes (when applicable). BONE
MARROW STAIN KIT FOR IRON Solution I - Potassium Ferrocyanide
(2%) RECOMMENDED PROCEDURE: Working Solution: Just prior to use, mix 12 ml of Sol. I with 36 ml of Sol. II. Fix preparation for 10 minutes in methanol. Stain for 10 minutes either by placing in a coplin jar or pouring directly over slide. Rinse slides with distilled or deionized water. Do not use tap water. Allow to drain and dry. Coverslip if desired. NOTE: The use of chemically clean glassware and plastic-tipped or paraffin-coated forceps is suggested. RESULTS: Hemosiderin and ferritin stain blue and is usually reported as either negative or 1+ to 5+. Iron in hemoglobin is unstained.
CONTENTS:
Potassium Ferrocyanide, HCl SIDEROCYTE
STAIN KIT Solution I - Potassium
Ferrocyanide (2%) RECOMMENDED PROCEDURE: Working Solution: Just prior to use, mix equal parts of Sol. I with Sol. II. Fix slide in methanol for 3 - 10 minutes and allow to dry. Flood slide with working solution for 2 - 5 minutes. Wash in distilled water and dry. Counterstain in Sol. III for 5 seconds. NOTE: The use of chemically clean glassware and plastic-tipped or paraffin-coated forceps is suggested. RESULTS: Siderotic granules stain bright blue.
CONTENTS: Potassium Ferrocyanide, HCl, Safranin O These
solutions are made from certified dyes (when applicable). PERIPHERAL
BLOOD & BONE MARROW STAIN KIT Solution I - Potassium
Ferrocyanide (4%) RECOMMENDED PROCEDURE: Working Solution: Just prior to use, mix equal parts of Solution I with Solution II. Fix slide in formalin vapor by placing in a closed staining jar containing a sponge or a piece of filter paper slightly moistened with formalin for 2 - 3 minutes. Slide is placed in the working solution that has been heated to 56oC or 132oF. Slide remains in working solution for 30 minutes and then removed and rinsed in tap water. Slide is then counterstained in Solution III - Basic Fuchsin solution for 5 minutes. Rinse slide in tap water, then ethanol and then again in water. Allow to air dry. NOTE: The use of chemically clean glassware and plastic-tipped or paraffin-coated forceps is suggested. RESULTS: Siderotic granules stain bright blue. Siderotic granules stain bright blue.
CONTENTS: Potassium Ferrocyanide, HCl, Safranin O Potassium Ferrocyanide, HCl, Basic Fuchsin, Phenol, Ethanol These
solutions are made from certified dyes (when applicable). SUDAN BLACK B STAIN KIT Solution I - Sudan Black B This technique may be substituted for the benzidine-peroxidase method used for the differential diagnosis of acute granulocytic leukemia, acute lymphocytic and acute myelo-monocytic leukemia. RECOMMENDED PROCEDURE: Working Solution: Prepare as needed by mixing 60 ml of Sol. I with 40 ml of Sol. II. Filter, using suction. Fixation: Place several pieces of filter paper in the bottom of a Coplin jar with screw-top lid. Moisten with 3 - 4 drops of 37% formaldehyde. Place air-dried smears in Coplin jar and cap tightly. Fix for 10 minutes. Wash in running water for 2 minutes. Place smears in working solution for 30 - 60 minutes. Wash in Sol. III for 30 - 60 seconds to remove excess stain. Wash in tap water for 2 minutes. Counterstain in Sol. IV for 10 minutes. Wash in tap water for 5 minutes. RESULTS: Lymphocytes and normoblasts are not stained with Sudan Black B. Normal granulocyte precursors from blast cells onward show increasing sudanophilia corresponding roughly to the granules present. Promyelocytes contain a few sudanophilic granules, while mature polymorphonuclear neutrophils contain large numbers of sudanophilic granules. Auer bodies are intensely sudanophilic.
CONTENTS: Sudan Black B, Ethanol, Buffers, Stabilizers, Hematoxylin, Potassium Alum, Mercuric Oxide. These
solutions are made from certified dyes (when applicable) FECAL STAIN KIT Solution I - Ethyl Alcohol (95%) Specifically for use in the microscopic examination of fecal material for fatty acids and neutral fats, as well as fecal leukocytes. RECOMMENDED PROCEDURE: Neutral Fats: Place a small aliquot of fecal suspension on a clean glass slide. Mix with 2 drops of Sol. I. Add and mix 2 drops of Sol. II. Coverslip and read microscopically. Fatty Acids: Place a small aliquot of fecal suspension on a clean, glass slide. Mix with 2 drops of Sol. III. Add and mix 2 drops of Sol. II. Coverslip and heat to boiling for a few seconds. Examine microscopically under high power while still warm. Fecal Leukocytes: Place a small aliquot of mucus or liquid stool on a clean, glass slide. Add 2 drops of Sol. IV and mix. Coverslip and allow to stand for 2 - 3 minutes before examining microscopically. NOTE: Fresh material should be used for optimum results. RESULTS: Neutral fats form yellow to pale orange refractile globules. Fatty acids appear as deep orange globules. To determine the presence of fecal leukocytes, examine stained specimen under low power. Differential counts (mononuclear or polymorphonuclear cells only) should be performed under high power.
CONTENTS:
Ethanol, Acetic Acid, Sudan IV, Methylene Blue87hese
solutions are made from certified dyes (when applicable). FECAL
LEUKOCYTE STAIN Specifically used in the microscopic identification and counting of fecal leukocytes. RECOMMENDED PROCEDURE: Place a small aliquot of mucus or liquid stool on a clean glass slide. Add 2 drops of Loeffler’s methylene blue and mix. Coverslip and allow to stand for 2 - 3 minutes before examining microscopically. NOTE: Fresh material should be used for optimum results. RESULTS: Fecal leukocytes stained with Loeffler’s methylene blue can be visualized under low power. Differential counts (mononuclear or polymorphonuclear cells only) should be performed under high power.
CONTENTS: Ethanol, Methylene Blue This solution is made from certified
dyes. METHYLENE BLUE
SOLUTIONS Solutions are used as a counterstain or for wet mount preparation. RECOMMENDED PROCEDURE: A small amount of feces is picked up with an applicator stick and thoroughly mixed with a large drop of methylene blue solution. The mixture is covered with a coverslip and sealed. The preparation should be examined within approximately 30 minutes. RESULTS: Trophozoite nuclei are stained blue with a lighter cytoplasm. Inclusions of the nuclei are also stained. After a time, the organisms become over-stained and are no longer identifiable.
CONTENTS: Methylene Blue These solutions are made from
certified dyes. SUDAN IV
(SCARLET RED) STAIN Solution I - Ethanol (95%) A simple qualitative screening procedure for the microscopic examination of fat in feces. RECOMMENDED PROCEDURE: Place a small aliquot of fecal suspension on a clean glass slide. Mix with 2 drops of Sol. I. Add and mix 2 drops of Sol. II. Coverslip and read microscopically. RESULTS: Neutral fats stain pale orange to red.
CONTENTS: Sudan IV, Ethanol These solutions are made
from certified dyes (when applicable). SUDAN IV
(SCARLET
RED) STAIN A simple qualitative screening procedure for the microscopic examination of fat in urine. RECOMMENDED PROCEDURE: Place a drop of urine from the surface of a centrifuged specimen or from centrifuged urine sediment on a clean glass slide, making two deposits. Add one drop of Sudan solution to one drop and coverslip both droplets individually. Look at unstained specimen before looking at stained droplet. The staining reaction can take a few minutes. RESULTS: Neutral fats stain pale orange to red. Neutral fats stain pale orange to red.
CONTENTS: Sudan IV, Ethanol This solution is made
from certified dye. in 2 sq. mm and add 5 zeros to obtain the number per ml. Vitality Staining: Mix a drop of semen with 2 drops Sol. IIC. Add 4 drops of Sol. IID, mix gently and prepare smears. Air dry. Staining For White Cells, Red Cells, Spermatozoa Morphology, etc.: Smear a drop of semen on a clean glass slide. Place in Sol. IIIA for 20 seconds, followed by Sol. IIIB for 20 seconds. SEMEN ANALYSIS KIT (UNAVAILABLE) Solution I - Semen Diluting Fluid This kit consists of a diluting fluid that renders spermatozoa immobile for easy chamber counting; a vitality stain that will easily differentiate between viable and non-viable spermatozoa; and a rapid stain and buffer solution requiring less than 60 seconds for the detection of pus cells (WBC’s, RBC’s, etc.). RECOMMENDED PROCEDURE: Sperm Count: Draw semen up to 0.5 mark (WBC pipette). Dilute to the 11.0 mark with Sol. I. Shake well. Charge hemacytometer and place in humidified chamber to avoid evaporation if unable to read immediately. Count the number of spermatazoa RESULTS: Total Count: 60,000,000 and higher. Vitality Stain: Dead, non-viable spermatozoa will stain red, while viable spermatozoa will be unstained (The Blom Technique). Stain For Cells, etc.: Similar to Wright Stain.
CONTENTS: Sodium Bicarbonate, Formalin (Neutral), Eosin, Nigrosin, Wright Stain, Methanol, Buffers These
solutions are made from certified dyes (when applicable). SEMEN ANALYSIS KIT (UNAVAILABLE) Solution I - Semen Diluting Fluid This kit consists of a diluting fluid that renders spermatozoa immobile for easy chamber counting; a vitality stain that will easily differentiate between viable and non-viable spermatozoa; and a rapid stain and buffer solution requiring less than 60 seconds for the detection of pus cells (WBC’s, RBC’s, etc.). RECOMMENDED PROCEDURE: Sperm Count: Draw semen up to 0.5 mark (WBC pipette). Dilute to the 11.0 mark with Sol. I. Shake well. Charge hemacytometer and place in humidified chamber to avoid evaporation if unable to read immediately. Count the number of spermatazoa in 2 sq. mm and add 5 zeros to obtain the number per ml. Vitality Staining: Mix a drop of semen with 2 drops Sol. IIA. Add 4 drops of Sol. IIB, mix gently and prepare smears. Fix by heat. Staining For White Cells, Red Cells, Spermatozoa Morphology, etc.: Smear a drop of semen on a clean glass slide. Place in Sol. IIIA for 20 seconds, followed by Sol. IIIB for 20 seconds. RESULTS: Total Count: 60,000,000 and higher. Vitality Stain: Dead, non-viable spermatozoa will stain red, while viable spermatozoa will be unstained (The Blom Technique). Stain For Cells, etc.: Similar to Wright Stain.
CONTENTS: Sodium Bicarbonate, Formalin (Neutral), Eosin, Nigrosin, Wright Stain, Methanol, Buffers These
solutions are made from certified dyes (when applicable). CSF
DILUTING FLUID A diluting fluid that hemolyzes all red cells and stains white cells for easy differentiation. RECOMMENDED PROCEDURE: Cell counts on spinal fluid should be made within 30 minutes after withdrawal of the specimen to avoid cell disintegration. Draw the specimen to the 1.0 mark (WBC pipette). Draw the diluting fluid to mark 11.0, producing a dilution of 1:10. Mix, discard one third, and place a drop on each side of the counting chamber. Count the cells in five squares. 1 sq. mm each (four corner squares and the central square), in both drops. Add the result of all 10 squares counted. Multiply by 10 to obtain the number of cells in 1 cmm. Use high power for a differential WBC count. NOTE: In some cases, to prevent possible coagulation of the spinal fluid due to the high protein-blood specimen, aqueous sodium citrate may be added in 1:100 dilution. Oxalates are unsatisfactory because the crystals interfere with counting.
CONTENTS: Glacial Acetic Acid, Crystal Violet This solution
is made from certified dyes. RETICULOCYTE
STAIN RECOMMENDED PROCEDURE: Generally, equal volumes of stain and blood are mixed together, allowed to stand 5 minutes and slides are made directly. No counterstaining is necessary. The proportion of blood to stain should be increased for low hemoglobins. May be used for finger sticks or anticoagulated blood. Retics Counted = Percent of Reticulocytes Count 1,000 red cells and record the number of reticulocytes present.
CONTENTS: New Methylene Blue, Potassium Oxalate This solution
is made from certified dyes. RETICULOCYTE
STAIN RECOMMENDED PROCEDURE: Generally, equal volumes of stain and blood are mixed together, allowed to stand 5 - 15 minutes and slides are made directly. To stain RBC’s and obtain darker reticulum, counterstain with Wright Stain (Product # 2000). The proportion of blood to stain should be increased for low hemoglobins. Retics Counted = Percent of
Reticulocytes Count 1,000 red cells and record the number of reticulocytes present.
CONTENTS: Brilliant Cresyl Blue, Sodium Citrate, Saline (NaCl) This solution
is made from certified dyes. SEMEN DILUTING FLUID RECOMMENDED PROCEDURE: Sperm Count: Draw semen up to 0.5 mark (WBC pipette) and dilute to 11.0 mark. Shake well, charge hemacytometer and place in humidified chamber to avoid evaporation if unable to read immediately. Count the number of spermatozoa in 2 sq. mm and add 5 zeros to obtain the number per ml.
CONTENTS: Sodium Bicarbonate, Formalin (Neutral) FOR IN VITRO DIAGNOSTIC USE ONLY. PLATELET DILUENT RECOMMENDED PROCEDURE: Draw blood up to the 0.5 mark (RBC pipette) and dilute to the 101 mark. Shake well approximately 1 minute. Expel fluid within the capillary end of the pipette. NOTE: Chamber and coverslip should be free of artifacts. Charge chamber, allow cells to settle (15 - 20 minutes) and count. Platelets Counted x Dilution(200)
x Depth(l0) = Platelets / cmm
CONTENTS: Ammonium Oxalate FOR IN VITRO DIAGNOSTIC USE ONLY. WBC DILUTING FLUID A modified solution which hemolyzes RBC’s and intensifies the cellular outline of WBC’s. RECOMMENDED PROCEDURE: Draw venous or capillary blood to the 0.5 mark (WBC pipette) and dilute to 11.0 mark. Charge chamber, allow cells to settle and count WBC’s in the four corner 1 sq. mm area. WBC’s Counted x
Dilution(20)xDepth(l0) = WBC’s / cmm
CONTENTS: Glacial Acetic Acid FOR IN VITRO DIAGNOSTIC USE ONLY. HEINZ BODY STAIN The detection of Heinz bodies is helpful in recognizing hemolytic anemia due to certain drugs and in detecting red cells of the primaquine-sensitive type. Heinz bodies are best stained supravitally. RECOMMENDED PROCEDURE: Mix a drop of blood with 8 drops of stain. Coverslip and examine under the oil immersion lens. RESULTS: Heinz bodies stain deep purple; Howell-Jolly bodies are always round, and Pappenheimer bodies stain darkly, almost black with a bluish tinge. Reticulocytes stain pale blue.
CONTENTS: Methyl Violet, Saline (NaCl) This solution
is made from certified dyes. RBC
DILUTING FLUID RECOMMENDED PROCEDURE: Draw blood to the 0.5 mark (Thoma pipette). Draw diluting fluid to the 101 mark. The resultant dilution is 1:200. Mix for 10 seconds. Expel fluid from capillary portion of pipette. Charge hemacytometer and allow cells to settle 1 - 3 minutes before counting. RBC’s Counted x Dilution(200) x Depth(l0) x Area(5) = RBC’s / cmm
CONTENTS: Sodium Sulfate, Sodium Chloride, Mercuric Chloride FOR IN VITRO DIAGNOSTIC USE ONLY. RBC DILUTING
FLUID RECOMMENDED PROCEDURE: Draw blood to the 0.5 mark (Thoma pipette). Draw diluting fluid to the 101 mark. The resulting dilution is 1:200. Mix for 2 - 3 minutes. Expel fluid from capillary portion of the pipette. Charge hemacytometer and allow cells to settle 1 - 3 minutes. RBC’s Counted x Dilution(200) x Depth(l0) x Area(5) = RBC’s/cmm
CONTENTS: Sodium Sulfate, Glacial Acetic Acid FOR IN VITRO DIAGNOSTIC USE ONLY. FETAL HEMOGLOBIN-DIFFERENTIAL STAIN KIT (UNAVAILABLE) Solution I - Citric Acid 0.lM This procedure, a modification of the Kleihauer Technique, is based on the resistance of fetal hemoglobin to acid conditions. By drying and fixation with ethanol, hemoglobin is precipitated within the red cells. Air-dried and fixed blood films are treated with a citric acid-phosphate buffer of low pH. Adult hemoglobin is rapidly eluted but HbF remains precipitated and is stained with eosin. RECOMMENDED PROCEDURE: Prepare thin (monolayer) blood films of capillary or fresh EDTA-anticoagulated blood and fix in 80% ethanol for 5 minutes. Rinse with water and allow to dry. Pipette 37.7 ml of Sol. I and 12.3 ml of Sol. II into a coplin jar and place in a 37oC water bath. When solution reaches 37oC, place fixed slide in coplin jar for 5 minutes. Move slides up and down a few times to ensure adequate elution of hemoglobin from cells. Remove slides and rinse with water. Stain with Sol. III for 1 - 4 minutes. Rinse with water. Stain with Sol. IV for 1 minute. Rinse with water and dry. Examine under low power for occasional eosin staining cells. Oil immersion is used for the pale ghosts cells that do not stain with eosin. RESULTS: RBC’s containing HbF stain brilliantly with eosin, while those without HbF appear as colorless "ghosts". The hematoxylin stains the WBC nuclei to prevent misinterpretation as eosin-positive cells.
CONTENTS: Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin These solutions are made from certified
dyes (when applicable). FETAL HEMOGLOBIN -
DIFFERENTIAL STAINING KIT (UNAVAILABLE) Modified Kleihauer Technique: Modified Clayton Technique: This product has been developed to employ a new modified Kleihauer procedure for fetal hemoglobin that does not require heat as in the old modified Kleihauer technique. This test can now be performed at room temperature. We recommend the use of hematoxylin for staining the WBC’s so as to avoid misidentification of eosin-positive cells. This is done with the new Kleihauer technique rather than the new Clayton technique. These procedures are based upon the resistance of fetal hemoglobin to acid conditions. Fixation of the smear allows hemoglobin to precipitate within the RBC. The Citric acid/Phosphate buffer solution eludes the adult hemoglobin. Fetal hemoglobin remains precipitated and stains bright red with eosin, making for easy differentiation. In the new and old Kleihauer procedure, WBC’s are counterstained with hematoxylin to avoid misidentification as eosin-positive cells. RECOMMENDED PROCEDURE: Prepare thin (monolayer) blood films of capillary or fresh EDTA-anticoagulated blood and fix in Sol. I for 5 minutes. Rinse with distilled water and allow to dry. Place slide in Sol.II or flood to elude 5 - 10 minutes. Rinse with distilled water. Stain with Sol. III for 1 - 4 minutes and rinse with distilled water. Stain with Sol. IV for 2 - 6 minutes. Rinse again in distilled water and allow time to dry.Prepare thin (monolayer) blood films of capillary or fresh EDTA-anticoagulated blood and fix in Sol. I for 5 minutes. Rinse with distilled water and allow to dry. Place slide in Sol.II or flood to elude 5 - 10 minutes. Rinse with distilled water. Stain with Sol. III for 1 - 4 minutes and rinse with distilled water. Stain with Sol. IV for 2 - 6 minutes. Rinse again in distilled water and allow time to dry. RESULTS: RBC’s containing HbF stain brilliantly with eosin, while those without HbF appear as colorless "ghosts". The hematoxylin stains the nuclei of the WBC’s blue to prevent misinterpretation as eosin-positive cells.
CONTENTS: Ethanol, Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin. Ethanol, Citric Acid, Disodium Phosphate, Hematoxylin, Sodium Iodate, Potassium Alum, Eosin. These solutions are made from certified
dyes (when applicable).
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