Hello friends,
Is it okay to use regular tissue culture/cell culture 96 well plates for measuring absorbance at 450nm? Although the wave length is in visible range I am bit skeptical about that and wanted confirmation from you guys. please share your thoughts..Thanks
I surely agree with your honest skepticism against the direct-use of photometer.
I must sadly and repeated say that Direct-utilization of photometer without purification is not quantitative due to violence against the Lambert-Beer's law (please see Fig.10 of the file; JCB Fucoidan transport). Similarly, notorious MTT method is also not quantitative at all, since this method uses photometer directly without purification.
The reliable viavility test should be performed as follows; i.e., 1) please scrape the cells by suitable cell-scraper, 2) wash the collected cells for 3-times by light centrifugation with PBS, 3) determine the amount of protein of cell-homogenate (ultra-sonic oscillation is convenient; please see files; J Chrom B rat BIN LIP Km and SEC column 300 A silica) of collected cells by the precize, quantitative, and quick HPLC-Surf-SEC protein-determination method (please see file; HPLC-Surf-SEC protein determination method).
Slow and steady wins the race.
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