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FORMALIN 10% NEUTRAL SALINE BUFFERED Intersep Product Code 1469 (250ml)
Formalin (10%) in 0.85% saline. This preserves the morphology of cysts better than that of 10% formalin solution. As with 10% formalin, trophozoites do not preserve well in formalin-saline and parasite morphology is not maintained adequately for a permanently stained faecal smear. It is recommended that 1 part of stool be mixed with 3 parts of formalin/formalin-saline preservative for the storage of bulk specimens.
SODIUM ACETATE-ACETIC FORMALIN ACID SOLUTION (SAF) Intersep Product Code 1461 (250ml)
Sodium acetate in Glacial acetic acid with 40% Formalin solution in distilled water. This preservative has the advantage of not containing mercuric chloride which both Schaudinns and PVA fixatives contain. This renders it less hazardous and more user friendly. This method preserves helminth eggs and larvae, protozoan cysts and trophozoites. This preservative can also be used for the formol-ethyl acetate concentration technique and for making permanent faecal smears.
Formol-ethyl acetate concentration methods can be performed on samples preserved by any of the above using Parasep
Parasitology
PARASITOLOGY STAINING METHODS
Temporary Staining Methods for Protozoa
Stains for wet preparations following concentration by the formol-ethyl acetate method.
1. ROMANOWSKY (FIELD A & B) STAINS 1.1 Modified Rapid Fields Stain Intersep Product Code 1482, 1483 (250ml)
This is a modification of Fields stain enabling rapid staining of fixed thin films of various clinical samples.This particular staining method is very useful for staining faecal smears, faecal exudate and duodenal aspirates. Method a) Make a thin film of faeces/exudate. Allow to air dry. b) Fix in methanol for 1 minute. c) Flood slide with 1 ml of Fields stain B (diluted 1:4 with distilled water) d) Immediately add an equal volume of undiluted Fields stain A, mix well and allow to stain for 1 minute. e) Rinse well in tap water and drain dry. Flagella, cilia and nuclei stain red and the cytoplasm stains blue.
1.2
Giemsa stain can also be used to stain films of unformed faeces, faecal exudate and duodenal aspirates. Method a) Make a thin film of faeces/exudate. Allow to air dry. b) Fix in methanol for 1 minute. c) Tip off the methanol and flood the slide with Giemsa stain diluted 1:10 with buffered distilled water. The diluted stain must be freshly prepared each time. d) Stain for 20-25 minutes. e) Run tap water on to the slide to float off the stain and to prevent deposition of precipitate on to the film. Allow to drain dry. f) Examine the film using the oil immersion objective. Flagella, cilia and nuclei stain red and the cytoplasm stains blue.
Parasitology
PARASITOLOGY PRESERVATION METHODS
A permanent Romanowsky stain (i.e. Giemsa or Rapid Fields stain) should be used for bloody diarrhoea and for semi-formed stools with blood and/or mucus. It provides information on: The exudate present. The presence of flagellate trophozoites. The presence of protozoa which are not readily detected or difficult to detect in the wet preparation eg. Dientamoeba fragilis and Blastocystis hominis. The presence of protozoa which are destroyed by the formol-ethyl acetate concentrate eg. Giardia lamblia.
Parasitology
PARASITOLOGY STAINING METHODS
Staining Iodine alcohol (solution B) 10 minutes. 70% ethanol 3-5 minutes. 80% ethanol 3-5 minutes. Trichrome stain 10 minutes. Acid alcohol (solution C) two immersions of one second each 95% ethanol two immersions 95 % ethanol 5 minutes. 95% ethanol 5 minutes. 100% ethanol 3 minutes. Xylene 5 minutes. Mount with a coverslip using DPX - do not allow the xylene to dry on the slide. Nuclei, chromidial bars, chromatin, red cells and bacteria stain red. Cytoplasm stains blue-green Background and yeasts stain green The spores of Microsporidia are very small - 1.0 x 0.5 m, oval in appearance and frequently exhibit a central band like structure flanked on either side by non-staining areas. Microsporidia spores stain pinkish red against a greenish background (Figure 14).
NB. the acetic acid does not dissolve the stain powder completely, it is quite normal to see a pink/red granular aggregate. Method a) Suspend unconcentrated stool in 10% formalin (1:3) b) Spread a small drop of stool suspension very thinly over 2/3 surface of a slide. c) Dry and fix in methanol for 5 minutes. d) Stain in Trichrome for 90 minutes at room temp. or stain for 10 minutes at 50C . e) Rinse in acid alcohol (4.5 ml glacial acetic acid in 95.5 ml of 90% ethanol) for 10 seconds. f) Rinse in 95 % ethanol until excess stain has washed off the slide. g) Dehydrate in 95% ethanol for 5 minutes. h) Dehydrate in 100% ethanol for 5 minutes. i) Clear in xylene for 5-10 minutes. j) Mount in DPX while still wet using a 22x40 coverslip and examine under oil immersion.
3. 4.
Parasitology
PARASITOLOGY PRESERVATION METHODS
All Intersep stains, reagents and fixatives are supplied prediluted ready for use.
Ordering Information
Product FIXATIVES
FORMALIN 10% NEUTRAL BUFFERED SAF FIXATIVE SCHAUDINNS FIXATIVE 10% FORMALIN FIXATIVE IN WATER BAYERS SOLUTION MERTHIOLATE-LODINE-FORMALIN (MIF) POLYVINYL ALCOHOL (PVA)
Pack Size
250ml 250ml 250ml 250ml 250ml 250ml 250ml
Ordering Code
1460 1461 1462 1463 1464 1465 1466
REAGENTS
MAYERS GLYCERIN/ALBUMIN PHYSIOLOGICAL SALINE TRITON X-100 SOLUTION ETHYL ACETATE ACETONE GRAMS DECOLOURISE 100ml 100ml 50ml 250ml 250ml 250ml 1470 1471 1472 1473 1475 1476
STAINS
EOSIN/SALINE MODIFIED ZIEHL-NEELSEN FUNGUQUAL (UVITEX 2B OR CALCOFLUOR) GRAM STAIN SAFRANIN NEUTRAL RED ACRIDINE ORANGE (ACETATE BUFFERED) AURAMINE PHENOL (LEMPERT) FlELD STAIN-SOLUTION A FIELD STAIN-SOLUTION B GIEMSA STAIN-RAPID IODINE-ALCOHOLIC LUGOLS IODINE-AQUEOUS IRON HAEMATOXYLIN-SOLUTION A IRON HAEMATOXYLIN-SOLUTION B TRICHROME FOR MICROSPORIDIA TRICHROME FOR PROTOZOA CRYSTAL VIOLET GRAMS FUCHSIN 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 250ml 1467 1468 1469 1491 1493 1494 1480 1481 1482 1483 1484 1485 1486 1487 1488 1489 1490 1492 1495
MOUNTING MEDIA
DPX MOUNTING MEDIA IMMERSION OlL MICROSCOPE SLIDES COVER GLASSES CARDBOARD SAMPLE SPOONS 100ml 50ml x100 x100 x50 1520 1521 1522 1523 1524