title: Identification of effective substrates for the direct analysis of lipids from cell lines using desorption electrospray ionization mass spectrometry.
abstract: BACKGROUND
  Various disease conditions, particularly tumours, can be understood easily by studying changes in the lipid profile of cells. While lipid profiles of tissues have been recorded by desorption electrospray ionization mass spectrometric (DESI-MS) imaging, there is paucity in standardized protocols for sample preparation involving cell cultures to generate reliable results. In this study, we report a method for the direct analysis of lipids from cultured cells by incorporating them onto Whatman 42 filter paper as a substrate for reliable DESI-MS analysis.
  
  
  METHODS
  The WERI-RB1 cell line was spotted on commonly used substrates for DESI-MS analysis, such as glass slides, Teflon coated glass slides, thin layer chromatography (TLC) plates, and Whatman 42 filter paper. A comparison of mass spectrometric images with two different lipids was made to understand the behaviour of different surfaces when the same sample was spotted on them. Relative intensities of different lipid peaks in the WERI-RB1 cell line were compared and relative lipid abundances were also compared across two different human retinoblastoma cell lines; WERI-RB1 and Y79.
  
  
  RESULTS
  The study demonstrates that good lipid signals can be obtained by DESI-MS when the cells are spotted on Whatman 42 filter paper. Tandem mass spectrometry was performed to identify the lipids as glycerophosphocholines (PC). Better lipid images from assembly of cells were obtained with distinct boundary when they were spotted on Whatman 42 filter paper than other surfaces.
  
  
  CONCLUSIONS
  We demonstrate the use of a simple substrate for reliable DESI-MS analysis of cultured cells. This method has the potential to understand various interactions of cells with other external agents. The current method would help in the application of DESI-MS for biology in general and medical sciences in particular.
authors.affiliation: Department of Chemistry, Indian Institute of Technology Madras, Chennai, 600036, India.
authors.forename: Amitava
authors.initials: A
authors.lastname: Srimany
authors.affiliation: Department of Nanobiotechnology, Vision Research Foundation, Sankara Nethralaya, Chennai, 600006, India.
authors.forename: Balasubramanyam
authors.initials: B
authors.lastname: Jayashree
authors.affiliation: Department of Nanobiotechnology, Vision Research Foundation, Sankara Nethralaya, Chennai, 600006, India.
authors.forename: Subramanian
authors.initials: S
authors.lastname: Krishnakumar
authors.affiliation: Department of Nanobiotechnology, Vision Research Foundation, Sankara Nethralaya, Chennai, 600006, India.
authors.forename: Sailaja
authors.initials: S
authors.lastname: Elchuri
authors.affiliation: Department of Chemistry, Indian Institute of Technology Madras, Chennai, 600036, India.
authors.forename: Thalappil
authors.initials: T
authors.lastname: Pradeep
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D008055:Lipids / Q000032:analysis*; D010627:Phenethylamines / Q000737:chemistry; D012175:Retinoblastoma / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D008055:Lipids; D010627:Phenethylamines; C029261:phenethylamine