PMID- 24123931
OWN - NLM
STAT- MEDLINE
DCOM- 20140930
LR  - 20191210
IS  - 2050-5701 (Electronic)
IS  - 2050-5698 (Linking)
VI  - 63
IP  - 1
DP  - 2014 Feb
TI  - Improvement of lateral resolution and extension of depth of field in two-photon 
      microscopy by a higher-order radially polarized beam.
PG  - 23-32
LID - 10.1093/jmicro/dft041 [doi]
AB  - The spatial resolution of laser scanning microscopes depends on the focal spot 
      size. As previously reported, we successfully improved the lateral spatial 
      resolution in confocal microscopy using liquid crystal devices (LCDs) to convert 
      a linearly polarized (LP) beam into a higher-order radially polarized (HRP) beam. 
      Taking advantage of the fact that those LCDs can be utilized at various 
      wavelengths, including near-infrared, we employed a near-infrared HRP beam to 
      improve the resolution in two-photon microscopy. Point-spread functions estimated 
      from fluorescent beads embedded in agarose gel showed that an HRP beam at 800-nm 
      excitation improved lateral resolution to 230 nm from 294 nm, which was obtained 
      using an LP beam at the same wavelength. Furthermore, at the glass-water 
      interface, the lateral resolution was considerably improved to 188 nm using the 
      HRP beam, whereas it degraded to 510 nm while using the LP beam. The HRP beams 
      visualized fine intracellular structures not only in fixed cells stained with 
      various dyes but also in living cells. Moreover, the HRP beam significantly 
      extended the depth of field, which facilitated obtaining in-focus images, 
      especially during time-lapse observations of living cells. These results indicate 
      that our method is applicable to various biological applications.
FAU - Ipponjima, Sari
AU  - Ipponjima S
AD  - Research Institute for Electronic Science, Hokkaido University, Kita 21 Nishi 10, 
      Kita-ku, Sapporo 001-0021, Japan.
FAU - Hibi, Terumasa
AU  - Hibi T
FAU - Kozawa, Yuichi
AU  - Kozawa Y
FAU - Horanai, Hibiki
AU  - Horanai H
FAU - Yokoyama, Hiroyuki
AU  - Yokoyama H
FAU - Sato, Shunichi
AU  - Sato S
FAU - Nemoto, Tomomi
AU  - Nemoto T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131011
PL  - England
TA  - Microscopy (Oxf)
JT  - Microscopy (Oxford, England)
JID - 101595834
SB  - IM
MH  - Animals
MH  - COS Cells
MH  - Chlorocebus aethiops
MH  - Microscopy, Confocal/*methods
MH  - Photons
OTO - NOTNLM
OT  - depth of field
OT  - liquid crystals
OT  - multi-photon microscopy
OT  - phase modulation
OT  - polarization
OT  - spatial resolution
EDAT- 2013/10/15 06:00
MHDA- 2014/10/01 06:00
CRDT- 2013/10/15 06:00
PHST- 2013/10/15 06:00 [entrez]
PHST- 2013/10/15 06:00 [pubmed]
PHST- 2014/10/01 06:00 [medline]
AID - dft041 [pii]
AID - 10.1093/jmicro/dft041 [doi]
PST - ppublish
SO  - Microscopy (Oxf). 2014 Feb;63(1):23-32. doi: 10.1093/jmicro/dft041. Epub 2013 Oct 
      11.