title: Silicon nitride as a versatile growth substrate for microspectroscopic imaging and mapping of individual cells.
abstract: 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.
authors.affiliation: School of Chemistry, The University of Sydney, NSW 2006, Australia.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Carter
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Rayner
authors.forename: Andrew I
authors.initials: AI
authors.lastname: McLeod
authors.forename: Lindsay E
authors.initials: LE
authors.lastname: Wu
authors.forename: Craig P
authors.initials: CP
authors.lastname: Marshall
authors.forename: Aviva
authors.initials: A
authors.lastname: Levina
authors.forename: Jade B
authors.initials: JB
authors.lastname: Aitken
authors.forename: Paul K
authors.initials: PK
authors.lastname: Witting
authors.forename: Barry
authors.initials: B
authors.lastname: Lai
authors.forename: Zhonghou
authors.initials: Z
authors.lastname: Cai
authors.forename: Stefan
authors.initials: S
authors.lastname: Vogt
authors.forename: Yao-Chang
authors.initials: YC
authors.lastname: Lee
authors.forename: Ching-Iue
authors.initials: CI
authors.lastname: Chen
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Tobin
authors.forename: Hugh H
authors.initials: HH
authors.lastname: Harris
authors.forename: Peter A
authors.initials: PA
authors.lastname: Lay
mesh_terms: 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
chemical_list: D017655:Silicon Compounds; C032734:silicon nitride