Cell smears

Cells containing Hb-F are densely stained with erythrosln and cells with Hb-A appear as ghost cells Intermediate cells are stained more or less pink. Reticulocytes with Hb-A sometimes resemble Intermediate cells and may also show some Intracellular granulation. Inclusion bodies are visible In eluted cells as compact particles of differing sizes. Figure 10 gives some examples. The method Is Ideally suited to demonstrate the presence of newborn red cells In the maternal circulation. The method Is also widely used for the evaluation of the distribution of Hb-F within red cells mainly to differentiate between the HPFH condition and the 3 or 36 thalassemias. Evaluation of F cell smears In such cases Is difficult the term "equal distribution" usually Indicates the presence of Hb-F In each red cell but not necessarily In the same amount. 

Figure 10, Red cell smears after the application of the add elution technique. A, artificial mixture of cells from newborn and adult. B, blood sample from a patient with Fanconts anemia (Hb-Fjto 14.6%).

Prior to beginning of IHC on formalin paraffin tissues more than 30 years ago, fresh cell smears or frozen tissue sections were used for immunofluorescence studies on tissues sections, and in a more limited way for immunperoxi-dase studies, now generally known as immunohistochemistry, prior to adaptation of the method to FFPE in 1974. The traditional viewpoint that... 

De Waele, M., Renmans, W., Segers, E., De Valck, V., Jochmans, K., and van Camp, B. (1989) An immunogold-silver staining method for detection of cell surface antigens in cell smears. J. Histochem. Cytochem. 37, 1855-1862. 

Air drying, usually with subsequent chemical post-fixation, is applicable for cell smears, cytospins and cryosections. Snap freezing (usually in liquid nitrogen) is routinely employed for tissue probes for subsequent cryosectioning. Chemical fixation is commonly carried out in aldehydes (e.g. formaldehyde) for tissue probes and cultured cell monolayers, and in acetone or alcohols (methanol or ethanol) for cryosections and cell preparations. 

This type of fixation is often used for cryosections, cell smears and cell monolyers, but can not be recommended for fixation of tissue blocks since acetone and alcohols, as opposed to aldehydes, penetrate tissue poorly. Cryosections, cell smears and cell monolyers after short (for 5 15 min) fixation in alcohols or acetone are usually air-dried (for 1 h or overnight), washed in buffered saline and directly subjected to the immunocytochemical analysis. 

Increased thickness of specimen Tissue sections should be cut 4-5 jm thick, and cell smears should be spread as thinly as possible. Cytospins should be only a monolayer in thickness. 

MICROWAVE HEAT-ASSISTED IMMUNOSTAINING OF CELL SMEARS... 

Thin, uniformly spread cell smears are prepared on glass slides and then rehydrated with normal saline for 3 min. This is followed by air-drying for 24 hr and fixation with 0.1% formal saline (1,000ml of normal saline and 2.5ml of 40% formalin) for 2-14 hr postfixation is accomplished with 95-100% ethanol for 10 min. The sections are heated in 10 mM citrate buffer (pH 6.0) in a microwave oven for 5 min at boiling and then allowed to remain in the hot solution for another 5 min before being removed for immunostaining. 

Finally, we wish to report the results for simulated cell smears. These experiments are being carried out to determine the best parameters for data acquisition, and to establish the variations of the cellular spectra from a homogeneous cell culture. Similar efforts have been undertaken before, where the cells were spin-deposited onto the microscope slides. However, for cultured cervical cancer (HeLa) cells, the resulting samples contained many cells that maintained their morphology in suspension quite well, and produced dried cells that were nearly spherical. Once dried, these cells could not be stained easily, and thus, their divisional activity could not be established after IR data acquisition. Furthermore, large spherical cells often gave spectral artifacts that made interpretation impossible.16... 

In this review, we demonstrate that excellent IR spectra from microscopic regions of cells and tissue can be collected. These spectra are extremely sensitive to variations in the biochemical composition of the pixels from which the spectra were acquired. Multivariate analyses of the spectra datasets of cells, cell smears and tissue sections produce pseudocolor maps in a totally unsupervised fashion that reproduce the histopathology of tissue sections and cytological features of cells and cell smears. 

This can be used in its acid form or as the sodium salt. The chromogens Fast Red TR and Fast Blue BB produce a bright red or blue end product, respectively. Both are soluble in alcoholic and other organic solvents, so aqueous mounting media must be used. Fast Red TR is preferred when staining cell smears. 

Fast Red Substrate Solution (recommended for cell smears)... 

Take small alicluots of cells to prepare cell smears on the slide for detection of viral antigens. Air-dry the cells, fix the cells in cold acetone for 10 min, air-dry, and then perform indirect immunofluorescent assay (see Appendix 1). 

Key words Cytocentrifuge, Cells, Smears, Touch preparations/imprints, DMSO, Fixation, Antibody penetration, Charged slides... 

Detection of different infectious agents, mainly viruses in tissue sections, cell smears and body fluids such as human papillomavirus (HPV), human herpesviruses including Epstein-Barr virus (EBV) and hepatitis C virus (HCV). ISH can detect latent viral genes and translation activity of infected cells as a criterion for latent infection. 

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