Fluorescent layers for characterization of sectioning microscopy with coverslipuncorrected and water immersion objectives

Handle URI:
http://hdl.handle.net/10754/563212
Title:
Fluorescent layers for characterization of sectioning microscopy with coverslipuncorrected and water immersion objectives
Authors:
Antonini, Andrea; Liberale, Carlo ( 0000-0002-5653-199X ) ; Fellin, Tommaso
Abstract:
We describe a new method to generate thin (thickness > 200 nm) and ultrathin (thickness < 200 nm) fluorescent layers to be used for microscope optical characterization. These layers are obtained by ultramicrotomy sectioning of fluorescent acrylic slides. This technique generates sub-resolution sheets with high fluorescence emission and uniform thickness, permitting to determine the z-response of different optical sectioning systems. Compared to the state of the art, the here proposed technique allows shorter and easier manufacturing procedure. Moreover, these fluorescent layers can be employed without protective coverslips, allowing the use of the Sectioned Imaging Property (SIP)-chart characterization method with coverslip-uncorrected objectives, water immersion objectives and micro-endoscopes. © 2014 Optical Society of America.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Label-Free Optical Microscopy for Biology Lab
Publisher:
The Optical Society
Journal:
Optics Express
Issue Date:
2014
DOI:
10.1364/OE.22.014293
Type:
Article
ISSN:
10944087
Sponsors:
We thank Roberta Ruffilli for her contribution to the preparation of the fluorescent layers, Mattia Pesce for his assistance in confocal measurements, Claudio Moretti for his help with two- photon measurements and Giuseppe Vicidomini for useful discussion. This work was supported by the Italian Institute of Technology "Interdisciplinary/Interdepartmental project" to TF and CL and grants from San Paolo "Programma in Neuroscienze", MIUR FIRB (RBAP11X42L), Telethon-Italy (GGP10138) and a FP7 DESIRE project to TF.
Appears in Collections:
Articles; Bioscience Program; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorAntonini, Andreaen
dc.contributor.authorLiberale, Carloen
dc.contributor.authorFellin, Tommasoen
dc.date.accessioned2015-08-03T11:38:16Zen
dc.date.available2015-08-03T11:38:16Zen
dc.date.issued2014en
dc.identifier.issn10944087en
dc.identifier.doi10.1364/OE.22.014293en
dc.identifier.urihttp://hdl.handle.net/10754/563212en
dc.description.abstractWe describe a new method to generate thin (thickness > 200 nm) and ultrathin (thickness < 200 nm) fluorescent layers to be used for microscope optical characterization. These layers are obtained by ultramicrotomy sectioning of fluorescent acrylic slides. This technique generates sub-resolution sheets with high fluorescence emission and uniform thickness, permitting to determine the z-response of different optical sectioning systems. Compared to the state of the art, the here proposed technique allows shorter and easier manufacturing procedure. Moreover, these fluorescent layers can be employed without protective coverslips, allowing the use of the Sectioned Imaging Property (SIP)-chart characterization method with coverslip-uncorrected objectives, water immersion objectives and micro-endoscopes. © 2014 Optical Society of America.en
dc.description.sponsorshipWe thank Roberta Ruffilli for her contribution to the preparation of the fluorescent layers, Mattia Pesce for his assistance in confocal measurements, Claudio Moretti for his help with two- photon measurements and Giuseppe Vicidomini for useful discussion. This work was supported by the Italian Institute of Technology "Interdisciplinary/Interdepartmental project" to TF and CL and grants from San Paolo "Programma in Neuroscienze", MIUR FIRB (RBAP11X42L), Telethon-Italy (GGP10138) and a FP7 DESIRE project to TF.en
dc.publisherThe Optical Societyen
dc.titleFluorescent layers for characterization of sectioning microscopy with coverslipuncorrected and water immersion objectivesen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentBioscience Programen
dc.contributor.departmentLabel-Free Optical Microscopy for Biology Laben
dc.identifier.journalOptics Expressen
dc.contributor.institutionDepartment of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia (IIT), Via Morego 30, 16163 Genova, Italyen
dc.contributor.institutionNanostructures Department, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genova, Italyen
kaust.authorLiberale, Carloen
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