Methodologies

Slide Preparation

Thin-spread glass slide preparations of bone marrow samples can be prepared in numerous ways, all of which have the aim to retain and evaluate marrow particles. Fat droplets and fragmented bone are likely to have adherent cellular material and thus be a target for morphologic evaluation.

An aspirate smear is similar in presentation as a peripheral blood smear.

	A drop of the specimen is placed 1 cm from the edge
	Place a second glass slide at 30º angle to the first slide & draw it back to make contact with sample.
	The sample is pushed toward the opposing side in one quick smooth stroke.
	Excess sample can be removed by tilting the glass slide onto gauze or pipetting the extraneous fluid.

Bone marrow aspiration and biopsy slides before staining
Obtained from: http://emedicine.medscape.com/article/207575-overview

Staining

Various stains can be used for different purposes

	Wright or Giemsa stain - enhance cytologic detail.
	Prussian blue - iron
	Myeloperoxidase/ Sudan Black B/ leukocyte alkaline phosphatase - categorization of acute myeloid leukemias
	Periodic acid-Schiff (PAS) stain - enhances depiction of cells (glycogen storage diseases)

Examination under microscope

Normal bone marrow
Note the presence of erythroid precursors and granulocytic precursors.
Obtained from: http://library.med.utah.edu/WebPath/HEMEHTML/HEME151.html

Presence of tumour cells in bone marrow smear from patient with Chronic Lymphocytic Leukemia
Obtained from: http://path.upmc.edu/cases/case141/micro.html

Leukemia
Marrow filled up with abnormal cells, displacing normal haematopoiesis
Obtained from: http://library.med.utah.edu/WebPath/HEMEHTML/HEME030.html

Normal Ranges of different cell types in bone marrow

Cell Type	Range %
Myeloblasts	0-2
Promyelocytes	2-5
Myelocytes (neutrophilic)	9-16
Metamyelocytes	7-23
Band forms	8-15
Neutrophils	4-10
Myelocytes (eosinophilic)	0-2
Band	0-2
Mature	0-3
Monocytes/macrophages	0-3
Basophils	0-1
Mast cells	0-2
Plasma cells	3-6

Obtained from: The Doctor's Doctor, http://www.thedoctorsdoctor.com/bodysites/bone_marrow.htm

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