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Ex vivo imaging of tumour-infiltrating T cells

SL Stefanie Lesch
VB Viktoria Blumenberg
SS Stefan Stoiber
AG Adrian Gottschlich
JO Justyna Ogonek
BC Bruno L. Cadilha
ZD Zahra Dantes
FR Felicitas Rataj
KD Klara Dorman
JL Johannes Lutz
CK Clara H. Karches
CH Constanze Heise
MK Mathias Kurzay
BL Benjamin M. Larimer
SG Simon Grassmann
MR Moritz Rapp
AN Alessia Nottebrock
SK Stephan Kruger
NT Nicholas Tokarew
PM Philipp Metzger
CH Christine Hoerth
MB Mohamed-Reda Benmebarek
DD Dario Dhoqina
RG Ruth Grünmeier
MS Matthias Seifert
AO Arman Oener
ÖU Öykü Umut
SJ Sandy Joaquina
LV Lene Vimeux
TT Thi Tran
TH Thomas Hank
TB Taisuke Baba
DH Duc Huynh
RM Remco TA. Megens
KJ Klaus-Peter Janssen
MJ Martin Jastroch
DL Daniel Lamp
SR Svenja Ruehland
MP Mauro Di Pilato
JP Jasper N. Pruessmann
MT Moritz Thomas
CM Carsten Marr
SO Steffen Ormanns
AR Anna Reischer
MH Michael Hristov
ET Eric Tartour
ED Emmanuel Donnadieu
SR Simon Rothenfusser
PD Peter Duewell
LK Lars M. König
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On day five after T cell transfer, the amount of CXCR6-GFP and GFP transduced T cells in tumour tissue was determined by 2-photon laser scanning microscopy (TPLSM). To distinguish intravascular and intratumoral T cells, blood vessels were stained by injecting mice with 3 μg anti-mouse CD31 antibody (eFluor450, clone 390, eBioScience) 30 min. before euthanasia. Imaging of tumour-infiltrating T cells was performed using a resonant scanning Leica SP5IIMP system equipped with a Spectra Physics MaiTai DeepSee Ti:Sa pulsed laser turned to 890 nm and a 20X NA 1.00 objective (Leica). Images with 1.5 to 2.0 μm spacing were acquired and processed using the Leica LAS X 3.1 software. The number of tumour-infiltrating T cells was quantified by counting of at least six representative areas per tumour.

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