Confocal light absorption and scattering spectroscopic microscopy monitors organelles in live cells with no exogenous labels.
This article reports the development of an optical imaging technique, confocal light absorption and scattering spectroscopic (CLASS) microscopy, capable of noninvasively determining the dimensions and other physical properties of single subcellular organelles. CLASS microscopy combines the principles of light-scattering spectroscopy (LSS) with confocal microscopy. LSS is an optical technique that relates the spectroscopic properties of light elastically scattered by small particles to their size, refractive index, and shape. The multispectral nature of LSS enables it to measure internal cell structures much smaller than the diffraction limit without damaging the cell or requiring exogenous markers, which could affect cell function. Scanning the confocal volume across the sample creates an image. CLASS microscopy approaches the accuracy of electron microscopy but is nondestructive and does not require the contrast agents common to optical microscopy. It provides unique capabilities to study functions of viable cells, which are beyond the capabilities of other techniques.
Itzkan
Irving
I
Department of Medicine, Beth Israel Deaconess Medical Center, Harvard University, Boston, MA 02215, USA.
Qiu
Le
L
Fang
Hui
H
Zaman
Munir M
MM
Vitkin
Edward
E
Ghiran
Ionita C
IC
Salahuddin
Saira
S
Modell
Mark
M
Andersson
Charlotte
C
Kimerer
Lauren M
LM
Cipolloni
Patsy B
PB
Lim
Kee-Hak
KH
Freedman
Steven D
SD
Bigio
Irving
I
Sachs
Benjamin P
BP
Hanlon
Eugene B
EB
Perelman
Lev T
LT
D002460:Cell Line; D002470:Cell Survival; D006801:Humans; D008853:Microscopy / Q000379:methods*; D015388:Organelles