Silicon nitride as a versatile growth substrate for microspectroscopic imaging and mapping of individual cells.
Herein is described a general sampling protocol that includes culture, differentiation and fixing of cells in their preferred morphology on the one sample substrate (Si(3)N(4)) to enable subsequent diverse modern microspectroscopic analyses. The protocol enables unprecedented correlated and complementary information on the intracellular biochemistry of metabolic processes, diseases and their treatment, which offers the opportunity to revolutionize our understanding of cell and tissue biology at a molecular level. The culture of adherent cells onto inexpensive Si(3)N(4) membranes allows microspectroscopic analyses across the electromagnetic spectrum, from hard X-ray fluorescence (both XRF and XANES), through to visible and fluorescence light microscopies, and infrared microspectroscopy without substrate interference. Adherent mammalian cell lines (3T3-L1 adipocytes and H9c2 cardiac myocytes) illustrate the in vitro application of these protocols. The cells adhered strongly to Si(3)N(4) membranes and visually displayed normal proliferative and phenotypic growth; more importantly, rapid alcohol fixation of cells did not affect their structural integrity for subsequent analyses.
Carter
Elizabeth A
EA
School of Chemistry, The University of Sydney, NSW 2006, Australia.
Rayner
Benjamin S
BS
McLeod
Andrew I
AI
Wu
Lindsay E
LE
Marshall
Craig P
CP
Levina
Aviva
A
Aitken
Jade B
JB
Witting
Paul K
PK
Lai
Barry
B
Cai
Zhonghou
Z
Vogt
Stefan
S
Lee
Yao-Chang
YC
Chen
Ching-Iue
CI
Tobin
Mark J
MJ
Harris
Hugh H
HH
Lay
Peter A
PA
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology*; D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D032383:Myocytes, Cardiac / Q000166:cytology*; D017655:Silicon Compounds / Q000737:chemistry / Q000378:metabolism*; D013052:Spectrometry, X-Ray Emission; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods; D017356:Synchrotrons
D017655:Silicon Compounds; C032734:silicon nitride