In vivo, realtime confocal imaging

J. V. Jester, P. M. Andrews, Walter M Petroll, M. A. Lemp, Harrison D Cavanagh

Research output: Contribution to journalArticle

95 Scopus citations

Abstract

We have adapted a tandem scanning confocal microscope for realtime, noninvasive imaging of cells under in vivo conditions. This form of in vivo confocal imaging relies on the optical sectioning abilities of the confocal microscope to obtain en face, sequential, reflected light images of cells at various depths, up to 1 mm, within opaque organs in living animals. Of major consideration in the design of an in vivo confocal microscope is maximizing the realtime detection of signals reflected from low contrast structures which can be affected by the microscope design, objective, and image detector systems. Using an in vivo confocal microscope design with a 20BioOptics surface contact objective we have obtained live cellular images from selected tissues including cornea, kidney, liver, adrenal, thyroid, epididymis, and muscle and connective tissue of rabbits and rats. Images were captured, digitized, and processed using a DAGE Mti low light level SIT camera coupled to a Gould IP9527 image processor. In vivo images were also compared with conventional bright field light and scanning electron microscopic images ofdead,fixed tissues. Overall, in vivo confocal imaging can provide remarkable detail of living cells comparable to that of conventional microscopic images ofdead,fixed, and stained tissue. A more unique feature of in vivo confocal imaging is the ability to study cellular structure and function sequentially over time in the same organ or tissue and represents a fundamentally new paradigm in microscopy. With continued refinements in the microscope, objective and detection system designs and their consequent improvements in lateral and axial resolution, in vivo confocal microscopy will enable us as observers to see what no one has been able to see before. Copyright1991 WileyLiss, Inc.

Original languageEnglish (US)
Pages (from-to)50-60
Number of pages11
JournalJournal of Electron Microscopy Technique
Volume18
Issue number1
DOIs
StatePublished - May 1991

Keywords

  • Confocal microscopy
  • Light microscopy
  • Scanning electron microscopy

ASJC Scopus subject areas

  • Anatomy

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