A Photomicrographic Method for the Detection of Oxygen Content in the Hemoglobin of the Red Blood Cell

Previous attempts at photographing the hemoglobin in the red blood cells have been confined to the making of two separate images of the same blood sample; one image is recorded using a wavelength band in the isobestic point region of the spectral distribution of oxygenated and reduced blood, and the other image is made using a wavelength band of radiation in the area of greatest difference in absorption between the two blood states. The method described here attempts to obtain a measurement of oxygenation by means of only one exposure using both regions of wavelengths simultaneously. The response is the density on film of the images of the two blood samples as recorded on color reversal film. A dilute blood sample was deoxygenated with nitrogen gas using a closed chamber. The blood solution was then transfered to a closed 100 micrometer depth chamber through which photomicrographs were made. Fully oxygenated blood was photographed In the same manner. Microdensitometric traces of the Images of individual oxygenated and reduced blood cells show that the meen density of the images of oxygenated hemoglobin in the cells was signifigantly greater than that of the reduced red blood cells