Subcellular and in-vivo Nano-Endoscopy.
Analysis of individual cells at the subcellular level is important for understanding diseases and accelerating drug discovery. Nanoscale endoscopes allow minimally invasive probing of individual cell interiors. Several such instruments have been presented previously, but they are either too complex to fabricate or require sophisticated external detectors because of low signal collection efficiency. Here we present a nanoendoscope that can locally excite fluorescence in labelled cell organelles and collect the emitted signal for spectral analysis. Finite Difference Time Domain (FDTD) simulations have shown that with an optimized nanoendoscope taper profile, the light emission and collection was localized within ~100 nm. This allows signal detection to be used for nano-photonic sensing of the proximity of fluorophores. Upon insertion into the individual organelles of living cells, the nanoendoscope was fabricated and resultant fluorescent signals collected. This included the signal collection from the nucleus of Acridine orange labelled human fibroblast cells, the nucleus of Hoechst stained live liver cells and the mitochondria of MitoTracker Red labelled MDA-MB-231 cells. The endoscope was also inserted into a live organism, the yellow fluorescent protein producing nematode Caenorhabditis elegans, and a fluorescent signal was collected. To our knowledge this is the first demonstration of in vivo, local fluorescence signal collection on the sub-organelle level.
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
Surya Venkatasekhar
SV
Cheemalapati
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
John
J
Winskas
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
Hao
H
Wang
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
Karthik
K
Konnaiyan
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
Arseny
A
Zhdanov
The Department of Cell Biology, Microbiology and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL 33620, USA.
Alison
A
Roth
The Department of Cell Biology, Microbiology and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL 33620, USA.
Swamy Rakesh
SR
Adapa
Department of Global Health, College of Public Health, University of South Florida, Tampa, FL 33647, USA.
Andrew
A
Deonarine
Department of Global Health, College of Public Health, University of South Florida, Tampa, FL 33647, USA.
Mark
M
Noble
Departments of Radiology and Integrated Mathematical Oncology, Moffitt Cancer Center, Tampa FL 33612, USA.
Tuhin
T
Das
Departments of Radiology and Integrated Mathematical Oncology, Moffitt Cancer Center, Tampa FL 33612, USA.
Robert
R
Gatenby
Department of Global Health, College of Public Health, University of South Florida, Tampa, FL 33647, USA.
Sandy D
SD
Westerheide
The Department of Cell Biology, Microbiology and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL 33620, USA.
Rays H Y
RH
Jiang
IBIS Lab, Department of Chemical and Biomedical Engineering, University of South Florida, Tampa, FL 33647, USA.
Anna
A
Pyayt
D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D045744:Cell Line, Tumor; D004724:Endoscopy; D006801:Humans; D008930:Mitochondria, Liver / Q000378:metabolism*; D057054:Molecular Imaging / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*
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