Quantification and calibration of images in fluorescence microscopy.
Fluorescence microscopy is a method widely used in life sciences to image biological processes in living and fixed cells or in fixed tissues. Quantification and calibration of images in fluorescence microscopy is notoriously difficult. We have developed a new methodology to prepare tissue "phantoms" that contain known amounts of (i) fluorophore, (ii) DNA, (iii) proteins, and (iv) DNA oligonucleotide standards. The basis of the phantoms is the ability of gelatin to act as a matrix for the conjugation of fluorophores as either a free-flowing liquid or a gelatinous solid depending on temperature (> or = 40 and < or = 4 degrees C).
Department of Neurosurgery, Methodist Hospital, Houston, TX 77030, USA.
David S
DS
Baskin
Marsha A
MA
Widmayer
Martyn A
MA
Sharpe
D000818:Animals; D002138:Calibration; D045744:Cell Line, Tumor; D004247:DNA / Q000032:analysis / Q000592:standards; D005456:Fluorescent Dyes / Q000032:analysis / Q000592:standards; D005780:Gelatin / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D009841:Oligonucleotides / Q000032:analysis / Q000592:standards; D019047:Phantoms, Imaging; D011506:Proteins / Q000032:analysis / Q000592:standards; D013552:Swine; D013696:Temperature
D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011506:Proteins; D005780:Gelatin; D004247:DNA