Abstract
Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical-aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25–40% for TPF images of subdiffraction fluorescent beads and 10–12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 129-135 |
| Number of pages | 7 |
| Journal | Journal of Microscopy |
| Volume | 271 |
| Issue number | 2 |
| DOIs | |
| State | Published - Aug 1 2018 |
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Keywords
- Multiphoton microscopy
- reflective objective
- second-harmonic generation
- two-photon fluorescence
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Histology
Cite this
Application of a reflective microscope objective for multiphoton microscopy. / Kabir, Mohammad M.; Choubal, Aakash M.; Toussaint, Kimani C.
In: Journal of Microscopy, Vol. 271, No. 2, 01.08.2018, p. 129-135.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Application of a reflective microscope objective for multiphoton microscopy
AU - Kabir, Mohammad M.
AU - Choubal, Aakash M.
AU - Toussaint, Kimani C
PY - 2018/8/1
Y1 - 2018/8/1
N2 - Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical-aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25–40% for TPF images of subdiffraction fluorescent beads and 10–12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications.
AB - Reflective objectives (ROs) mitigate chromatic aberration across a broad wavelength range. Yet, a systematic performance characterisation of ROs has not been done. In this paper, we compare the performance of a 0.5 numerical-aperture (NA) reflective objective (RO) with a 0.55 NA standard glass objective (SO), using two-photon fluorescence (TPF) and second-harmonic generation (SHG). For experiments spanning ∼1 octave in the visible and NIR wavelengths, the SO leads to defocusing errors of 25–40% for TPF images of subdiffraction fluorescent beads and 10–12% for SHG images of collagen fibres. The corresponding error for the RO is ∼4% for both imaging modalities. This work emphasises the potential utility of ROs for multimodal multiphoton microscopy applications.
KW - Multiphoton microscopy
KW - reflective objective
KW - second-harmonic generation
KW - two-photon fluorescence
UR - http://www.scopus.com/inward/record.url?scp=85051431714&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85051431714&partnerID=8YFLogxK
U2 - 10.1111/jmi.12702
DO - 10.1111/jmi.12702
M3 - Article
VL - 271
SP - 129
EP - 135
JO - Journal of Microscopy
JF - Journal of Microscopy
SN - 0022-2720
IS - 2
ER -