Endoscopy cornerMultiphoton Imaging Can Be Used for Microscopic Examination of Intact Human Gastrointestinal Mucosa Ex Vivo
Section snippets
Acquisition of Human Samples
Patients presenting to the Yale Gastrointestinal Procedure Center for elective outpatient endoscopy were recruited to participate in this study, which was approved by the Yale University Human Investigations Committee. Written informed consent was obtained before study participation. Inclusion criteria included age older than 18 years and the ability to provide informed consent. Exclusion criteria included a known or suspected bleeding disorder, international normalized ratio exceeding 1.4,
Optimal Multiphoton Excitation Wavelength for Mucosa
To determine the optimal multiphoton excitation wavelength for tissue autofluorescence and collagen SHG in gastrointestinal tissue, a femtosecond-pulsed, mode-locked Ti:S laser was used to examine fresh untreated segments of rat colon. The laser was tuned across a range of wavelengths between 715 and 790 nm. Fluorescence emission was collected in the 410- to 490-nm range, and the mean fluorescence intensity across the field of view was calculated. Figure 1 shows the relationship between
Discussion
Initial reports using confocal laser scanning microscopy to examine gastrointestinal mucosa ex vivo showed limited image detail in specimens that were not treated with fluorescent dyes.15, 16 Our results show that multiphoton laser microscopy provides the ability to detect cellular and subcellular details in unfixed, unstained gastrointestinal mucosa, with image quality that is superior to confocal imaging with or without fluorescent staining. Throughout the gastrointestinal tract, mucosal
References (34)
- et al.
Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo
Gastroenterology
(2004) - et al.
Application of multiline two-photon microscopy to functional in vivo imaging
J Neurosci Meth
(2006) - et al.
New developments in multiphoton microscopy
Curr Opin Neurobiol
(2002) - et al.
Interpreting second-harmonic generation images of collagen I fibrils
Biophys J
(2005) - et al.
In vivo histology of Barrett’s esophagus and associated neoplasias by confocal laser endomicroscopy
Clin Gastroenterol Hepatol
(2006) - et al.
Chromoscopy-guided endomicroscopy increases the diagnostic yield of intraepithelial neoplasia in ulcerative colitis
Gastroenterology
(2007) - et al.
Laser-induced fluorescence spectroscopy of human colonic mucosaDetection of adenomatous transformation
Gastroenterology
(1990) - et al.
Flexible transgastric peritoneoscopy: a novel approach to diagnostic and therapeutic interventions in the peritoneal cavity
Gastrointest Endosc
(2004) - et al.
A miniature head-mounted two-photon microscope: high resolution brain imaging in freely moving animals
Neuron
(2001) - et al.
In vivo imaging of the dendritic arbors of layer V pyramidal cells in the cerebral cortex using a laser scanning microscope with a stick-type objective lens
Neurosci Lett
(2006)
Confocal laser endomicroscopy: technical status and current indications
Endoscopy
Adenomatous and hyperplastic polyps cannot be reliably distinguished by their appearance through the fiberoptic sigmoidoscope
Dig Dis Sci
Nonlinear magic: multiphoton microscopy in the biosciences
Nat Biotechnol
Deep tissue two-photon microscopy
Nat Methods
Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation
Proc Natl Acad Sci U S A
Two-photon 3-D mapping of ex vivo human skin endogenous fluorescence species based on fluorescence emission spectra
J Biomed Opt
Intravital multiphoton microscopy of dynamic renal processes
Am J Physiol
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Supported by National Institute of Health grants P30 DK34989 and R01 DK45710 (M.H.N.) and R01 CA116583, P41 EB01976, and P41 RR04224 (W.R.Z.). Also supported by a pilot grant from the Pentax Corporation (Montvale, NJ) to support the cost of processing biopsy specimens for H&E staining.