Spatially Co-registered Wide-field Nonlinear Optical Imaging of Living and Complex Biosystems in a Total Internal Reflection Geometry

The Analyst ◽  
2021 ◽  
Author(s):  
Uvinduni I. Premadasa ◽  
Amber N. Bible ◽  
Jennifer L Morrell-Falvey ◽  
Benjamin Doughty ◽  
Yingzhong Ma
Keyword(s):  
Total Internal Reflection ◽  
Nonlinear Optical ◽  
Internal Reflection ◽  
Wide Field ◽  
Excitation Scheme ◽  
Surface Sensitivity ◽  
Nonlinear Optical Imaging ◽  
Reflection Geometry ◽  
Enhanced Surface ◽  
Field Imaging

Nonlinear optical microscopy that leverages an objective based total internal reflection (TIR) excitation scheme is an attractive means for rapid, wide-field imaging with enhanced surface sensitivity. Through select combinations of...

Measurement of Ca2+ signaling dynamics in exocrine cells with total internal reflection microscopy

2006 ◽  
Vol 291 (1) ◽  
pp. G146-G155 ◽  
Author(s):  
Jong Hak Won ◽  
David I. Yule
Keyword(s):  
Plasma Membrane ◽  
Total Internal Reflection ◽  
Acinar Cells ◽  
Internal Reflection ◽  
Pancreatic Acinar Cells ◽  
Wide Field ◽  
Exocrine Cells ◽  
Parotid Acinar Cells ◽  
Signaling Dynamics ◽  
Total Internal Reflection Microscopy

In nonexcitable cells, such as exocrine cells from the pancreas and salivary glands, agonist-stimulated Ca2+ signals consist of both Ca2+ release and Ca2+ influx. We have investigated the contribution of these processes to membrane-localized Ca2+ signals in pancreatic and parotid acinar cells using total internal reflection fluorescence (TIRF) microscopy (TIRFM). This technique allows imaging with unsurpassed resolution in a limited zone at the interface of the plasma membrane and the coverslip. In TIRFM mode, physiological agonist stimulation resulted in Ca2+ oscillations in both pancreas and parotid with qualitatively similar characteristics to those reported using conventional wide-field microscopy (WFM). Because local Ca2+ release in the TIRF zone would be expected to saturate the Ca2+ indicator (Fluo-4), these data suggest that Ca2+ release is occurring some distance from the area subjected to the measurement. When acini were stimulated with supermaximal concentrations of agonists, an initial peak, largely due to Ca2+ release, followed by a substantial, maintained plateau phase indicative of Ca2+ entry, was observed. The contribution of Ca2+ influx and Ca2+ release in isolation to these near-plasma membrane Ca2+ signals was investigated by using a Ca2+ readmission protocol. In the absence of extracellular Ca2+, the profile and magnitude of the initial Ca2+ release following stimulation with maximal concentrations of agonist or after SERCA pump inhibition were similar to those obtained with WFM in both pancreas and parotid acini. In contrast, when Ca2+ influx was isolated by subsequent Ca2+ readmission, the Ca2+ signals evoked were more robust than those measured with WFM. Furthermore, in parotid acinar cells, Ca2+ readdition often resulted in the apparent saturation of Fluo-4 but not of the low-affinity dye Fluo-4-FF. Interestingly, Ca2+ influx as measured by this protocol in parotid acinar cells was substantially greater than that initiated in pancreatic acinar cells. Indeed, robust Ca2+ influx was observed in parotid acinar cells even at low physiological concentrations of agonist. These data indicate that TIRFM is a useful tool to monitor agonist-stimulated near-membrane Ca2+ signals mediated by Ca2+ influx in exocrine acinar cells. In addition, TIRFM reveals that the extent of Ca2+ influx in parotid acinar cells is greater than pancreatic acinar cells when compared using identical methodologies.

scholarly journals Giant Terahertz-Wave Absorption by Monolayer Graphene in a Total Internal Reflection Geometry

ACS Photonics ◽  
2016 ◽  
Vol 4 (1) ◽  
pp. 121-126 ◽  
Author(s):  
Yoichi Harada ◽  
Muhammad Shoufie Ukhtary ◽  
Minjie Wang ◽  
Sanjay K. Srinivasan ◽  
Eddwi H. Hasdeo ◽  
...  
Keyword(s):  
Total Internal Reflection ◽  
Terahertz Wave ◽  
Internal Reflection ◽  
Monolayer Graphene ◽  
Wave Absorption ◽  
Reflection Geometry

scholarly journals Extrinsic Absorption Pathways in Vanadium‐Doped SiC Measured Using a Total Internal Reflection Geometry

2020 ◽  
Vol 217 (20) ◽  
pp. 2000315
Author(s):  
Noah J. Kramer ◽  
Lars F. Voss ◽  
Adam M. Conway ◽  
Paulius V. Grivickas ◽  
Mihail Bora ◽  
...  
Keyword(s):  
Total Internal Reflection ◽  
Internal Reflection ◽  
Reflection Geometry ◽  
Absorption Pathways ◽  
Extrinsic Absorption

scholarly journals Total internal reflection enabled wide-field coherent anti-Stokes Raman scattering microscopy

2020 ◽  
Vol 45 (11) ◽  
pp. 3087
Author(s):  
Benjamin Doughty ◽  
Uvinduni I. Premadasa ◽  
John F. Cahill ◽  
Amber B. Webb ◽  
Jennifer L. Morrell-Falvey ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Wide Field ◽  
Anti Stokes
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