Determination of cell concentrations using haemocytometer according to Fuchs Rosenthal and
Burker [Internet].
Standard procedures were employed for total and differential cell numbers evaluation: for valuation of leukocytes, sediment cells from BALF were suspended in PBS treated with Turk reagents and calculated in optical microscopy in a
Burker chamber.
The enumeration was performed by using
Burker chamber and sperm count was calculated using weight of testis and dilution volumes.
KBr-IR and [sup.13]C, [sup.1]H NMR spectra were obtained by JASCO FT-IR-4100 spectrophotometer (400-4000 [cm.sup.-1]) and
Burker 500 MHz, respectively.
The protocol was designed to validate an automated method for MSC counting by "NucleoCounter[R]" system (ChemoMetec, Allerod, Denmark) in terms of accuracy, precision, and linearity in comparison to the manual cell count method by the hemocytometer (
Burker chamber).
The DNA content was expressed as [micro]g DNA/[10.sup.6] cells counted, in each well, using a
Burker chamber and trypan blue.
1H-NMR spectra were recorded in CDCl3 on
Burker spectrometer, operating at 400 MHz at 25 AdegC.
Cell concentrations, in number per milliliter, were determined by use of a hemocytometer (
Burker) by counting 150-300 cells as described elsewhere (29).