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DTSTART:20170326T030000
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DTSTART:20161030T020000
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DTSTAMP:20260428T224327Z
UID:58bd32a703c04556423447@ist.ac.at
DTSTART:20170308T160000
DTEND:20170308T170000
DESCRIPTION:Speaker: Wiebke Jahr\nhosted by Johann Georg Danzl\nAbstract: A
  major goal in biological imaging is to visualize interactions of differen
 t tissues\, often fluorescently labeled\, during dynamic processes. Only a
  few of these labels fit into the available spectral range without overlap
 \, but can be separated computationally if the full spectrum of every sing
 le pixel is known. In medical imaging\, hyperspectral techniques show prom
 ise to identify different tissue types without any staining. Yet\, microsc
 opists still commonly acquire spectral information either with filters\, t
 hus integrating over a few broad bands only\, or point-wise\, dispersing t
 he spectra onto a multichannel detector\, which is inherently slow. Light 
 sheet fluorescence microscopy (LSFM) and optical projection tomography (OP
 T) are two techniques to acquire 3D microscopic data fast\, photon-efficie
 ntly and gently on the specimen. LSFM works in fluorescence mode and OPT i
 n transmission. Both are based on a fast widefield detection scheme where 
 a 2D detector records the spatial information but leaves no room to acquir
 e dispersed spectra. Hyperspectral imaging had not yet been demonstrated f
 or either technique. I present a line-scanning hyperspectral LSFM and an e
 xcitation scanning OPT to acquire 5D data and show how the performance of 
 both setups can be optimized to minimize acquisition times without sacrifi
 cing image contrast\, spatial or spectral information. We implemented and 
 assessed different evaluation pipelines to classify and unmix relevant fea
 tures from the hyperspectral data. We demonstrated the efficiency of the w
 orkflow by acquiring up to five fluorescent markers and the autofluorescen
 ce in zebrafish and fruit fly embryos on the hyperspectral LSFM. Both conc
 entration maps and spectra for each of the fluorophores were extracted usi
 ng linear unmixing and spectral phasor analysis. The same methods were app
 lied to investigate the transmission data acquired on the spectral OPT\, w
 here we found evidence that OPT image formation is governed by refraction\
 , whereas scattering and absorption only play a minor role.\n
LOCATION:Mondi Seminar Room 1\, Central Building\, ISTA
ORGANIZER:rsix@ist.ac.at
SUMMARY:Wiebke Jahr: Hyperspectral three-dimensional widefield microscopy i
 n living zebrafish and fruit fly embryos
URL:https://talks-calendar.ista.ac.at/events/357
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