Nanolive SA invites you to its first lunch seminar given by the group of
Professor Tsutomu Masujima from RIKEN, Japan
DATE: Thursday June 1st
TIME: 12:15 – 13:00
LOCATION: Nanolive SA, Batiment L Innovation Park, EPFL
PIZZA & SOFT DRIKS WILL BE PROVIDED
RSVP: here
Absolute quantitation of a single cancer cell by live single cell mass spectrometry coupled with 3D holography and tomography.
(Quantitative Biology Center (QBiC), RIKEN.1, Faculty of Pharmacy, Misr International University 2, Nanolive SA 3)
A. Ali,1,2, , Y.Abouleila,1,2, Sara AMER, 1,2 Rie FURUSHIMA, 1 Samy EMARA, 2 Sebastien EQUIS, 3 Yann COTTE,3 and Tsutomu MASUJIMA1
Keywords: Absolute Quantitation, Live Single Cell Mass Spectrometry, 3D Holography and tomorgraphy
A single HepG2 cell was visualized in three-dimensional 3D holographic and tomographic microscope. While the cell is alive, it was digitally stained according to the values of each component’s refractive index. A micromanipulator was connected to the body of the microscope by a common base plate, a 1um internal diameter Nanospray tip was attached to the micromanipulator which was used to suck out the content of the single HepG2 cell that was monitored in 3D. Volume analysis was carried out by means of 3D holographic image analysis and the volume of cytoplasm, nucleus, endoplasmic reticulum, and a single granule was calculated. Moreover, a 3D image was taken of the cell before and after suction of the cytoplasm and nucleus to determine the exact volume by the Nanospray tips for later quantitation. Target molecules were chosen per their specific site in the cell and Nanospray tips were used to take samples from the cytoplasm and the nucleus. Each concentration was calculated only in the volume taken from the cell.
Cytoplasm specific molecules (N-Acetyl putrescine and Methionine Sulfoxide) were successfully quantified to be 6.0 zmole/1.16 pL and 5.9zmol/1.16 pL respectively. Moreover, Nucleus specific 3’-O-Methyguanosine concentration was determined to be 28.7 zmole/0.96pL.
References
1) G. J. Brakenhoff, P. Blom, and P. Barends, , J. Microscopy, 1979, 117, 219
2) Y. Hiraoka, T. Shimi, and T. Haraguchi, Cell Struct. Funct. 2002, 27, 367.
3) Y. Cotte, F. Toy, P. Jourdain, N. Pavillon, D. Boss, P. Magistretti, P. Marquet, and C. Depeursinge, Nature Photonics, 2013, 7, 113.
4) N. Tsuyama, H. Mizuno, E. Tokunaga, and T. Masujima, Anal. Sci., 2008, 24, 559.
5) H. Mizuno, N. Tsuyama, T. Harada, and T. Masujima, J. Mass Spectrum., 2008, 43, 1692.
6) T. Masujima, Anal. Sci., 2009, 25, 953.
7) T. Fujii, S. Matsuda, M. L. Tejedor, T. Esaki, I. Sakane, H. Mizuno, N. Tsuyama, and T. Masujima, Nat. Protoc., 2015, 10, 1445.
8) C. Schneider, W. Rasband, and K. Eliceiri, Nature methods, 2012, 9, 671.
9) J. Schindelin, I. Arganda-Carreras, and E. Frise et al., Nature methods, 2012, 9, 676.
10) S. Bolte and F. P. Cordelières, , J. Microscopy, 2006, 224,213.
