Live cell refractometry using Hilbert phase microscopy and confocal reflectance microscopy.
Quantitative chemical analysis has served as a useful tool for understanding cellular metabolisms in biology. Among many physical properties used in chemical analysis, refractive index in particular has provided molecular concentration that is an important indicator for biological activities. In this report, we present a method of extracting full-field refractive index maps of live cells in their native states. We first record full-field optical thickness maps of living cells by Hilbert phase microscopy and then acquire physical thickness maps of the same cells using a custom-built confocal reflectance microscope. Full-field and axially averaged refractive index maps are acquired from the ratio of optical thickness to physical thickness. The accuracy of the axially averaged index measurement is 0.002. This approach can provide novel biological assays of label-free living cells in situ.
G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Niyom
N
Lue
Wonshik
W
Choi
Gabriel
G
Popescu
Zahid
Z
Yaqoob
Kamran
K
Badizadegan
Ramachandra R
RR
Dasari
Michael S
MS
Feld
D048430:Cell Shape; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008858:Microscopy, Phase-Contrast; D012031:Refractometry