HEMATOLOGICAL STAINING TECHNIQUES

TAREMWA Ivan MHTM, 2014.*

*

Blood Composition*

Diagnostic trends*

Any challenges with these slides?*

Forensic Characterization of BloodHemoglobin possesses peroxidase like activity which when mixed with phenolphthalein and hydrogen peroxide it will cause the formation of a deep pink color

Once blood is found it must be determined to be humanPrecipitin test is the standard testReagents are available to type if blood is dog,cat or othersA positive test is a cloudy ring or band at the point where the two liquids meetOnly a small amount of sample is requiredPrecipitin test is very sensitive and can test positive on a sample as old as 10 years

*

Blood is examined in a smear

Smears are stained using Romanowsky stains; Giemsa, Jenner, Wright, Field and Leishman.*

Reticulocyte stains and CountBetter and more reliable results are obtained with New methylene blue, (? different from methylene blue) Brilliant cresyl blue and Azure B. - Reticulofilamentous material.MethodDeliver 2 or 3 drops of the dye solution in a tube- Supravital staining. Add 24 volumes of the patients EDTA-anticoagulated blood to the dye solution and mix. Keep the mixture at 37 0C for 1520 min. Resuspend the red cells by gentle mixing and make films on glass slides the usual way. When dry, examine the films without fixing or counterstaining.*

*

Place a small drop of blood from the frosted end of a slide.The end of a second slide is used to spread the blood drop out on the first slide.

Pull the end of the slide through the blood drop toward you and then push in the opposite direction forcing the blood in your direction.

*

Staining the blood smear: The smear should be stained as soon as the slide dries.

If the slide cannot be stained immediately, it can be immersed in methanol (fixed) and stained later.

*

Staining blood and BM filmsRomanowsky stains are used universally for routine staining of blood films; with satisfactory results.

These offers subtle distinctions in shades of staining between nucleus & cytoplasmic contents.

*

Color patterns of blood cells

*

Cell Morphology*LymphocyteSegmented NeutrophilPlateletrbc

*neutrophileosinophilbasophilsmall lymphocytemonocyte

The Peripheral Smear*

SchistocytesHemolytic Anemia, BurnsSpherocytesAIHA, Hereditary SpherocytosisTarget cellsAlcoholics, HbpathiesSideroblastsMyelodysplasia, AlcoholicsTeardrop cellsMyelofibrosis/Myeloid metaplasia, ThalassemiaBurr cellsUremiaHowell Jolly bodiesSplenectomy, Functional aspleniaHypersegmented PMNMegaloblastic Anemia

What are these?What stains were used?*

Blood smear in a 43 year old man with history of a motor vehicle accident 12 years ago.

What is this?What may have happened?*

Case 1: Liz, a PG, has just undergone C/S . She reports surgery in childhood and she shows signs of infections. Her CBC is normal [as below]:

Wbc 1.7 Hgb 8.9 g/dLPlatelets 109,000MCV 109Segs 52%Lymphs 40%Mono 5%Eos 2%Metamyelocytes 1%

Considering the PBF, what definitive diagnosis can you give?

Would this differ from the definitive diagnosis without the PBF?

*Additional tests

Questions?*

**