+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Application of multiline two-photon microscopy to functional in vivo imaging

, : Application of multiline two-photon microscopy to functional in vivo imaging. Journal of Neuroscience Methods 151(2): 276-286

High spatial resolution and low risks of photodamage make two-photon laser-scanning microscopy (TPLSM) the method of choice for biological imaging. However, the study of functional dynamics such as neuronal calcium regulation often also requires a high temporal resolution. Hitherto, acquisition speed is usually increased by line scanning, which restricts spatial resolution to structures along a single axis. To overcome this gap between high spatial and high temporal resolution we performed TPLSM with a beam multiplexer to generate multiple laser foci inside the sample. By detecting the fluorescence emitted from these laser foci with an electron-multiplying camera, it was possible to perform multiple simultaneous linescans. In addition to multiline scanning, the array of up to 64 laser beams could also be used in x-y scan mode to collect entire images at high frame rates. To evaluate the applicability of multiline TPLSM to functional in vivo imaging, calcium signals were monitored in visual motionsensitive neurons in the brain of flies. The capacity of our method to simultaneously acquire signals at different cellular locations is exemplified by measurements at branched neurites and 'spine'-like structures. Calcium dynamics depended on branch size, but 'spines' did not systematically differ from their 'parent neurites'. The spatial resolution of our setup was critically evaluated by comparing it to confocal microscopy and the negative effect of scattering of emission light during image detection was assessed directly by running the setup in both imaging and point-scanning mode. (c) 2005 Elsevier B.V. All rights reserved.


PMID: 16442636

DOI: 10.1016/j.jneumeth.2005.12.003

Other references

Ogden, J.E.; Fantes, P.A., 1986: Isolation of a novel type of mutation in the mitotic control of Schizosaccharomyces pombe whose phenotypic expression is dependent on the genetic background and nutritional environment. The major cell cycle control in the fission yeast Schizosaccharomyces pombe acts at entry to mitosis, and involves three previously identified genes cdc2, cdc25 and wee1. The presence of a wee1 mutation phenotypically suppresses cdc25 mutations. T...

Shannon, Derek R., 1995: Annual report of the council for the year ended 31 July 1995. Transactions of the Natural History Society of Northumbria. November; 566: 393-420

Henriques, J.F.; Martins, D.R.; Pinzan, A., 1979: The cervical headgear action in the mixed dentition on maxilla, mandible and teeth in class II, division 1, malocclusions--a cephalometric study. Ortodontia 12(2): 76-86

White, M.K.; Kaminski, R.; Young, W-Bin.; Roehm, P.C.; Khalili, K., 2017: CRISPR Editing Technology in Biological and Biomedical Investigation. The CRISPR or clustered regularly interspaced short palindromic repeats system is currently the most advanced approach to genome editing and is notable for providing an unprecedented degree of specificity, effectiveness, and versatility in genetic...

Sasai, Y., 1968: Histochemical demonstration of glycogen synthetase in the normal human epidermis. Tohoku Journal of Experimental Medicine 96(1): 75-80

Lin, L-Fu.; Huang, P-Teh., 2012: Linear endoscopic ultrasound for clinically suspected bile duct stones. Radial endoscopic ultrasound (EUS) is a less invasive imaging modality used to evaluate common bile duct stones (CBDS) and is as accurate as endoscopic retrograde cholangiography (ERCP). Patients classified as high risk for suspected CBDS were fou...

Martinez, R.; Esteller, M., 2010: The DNA methylome of glioblastoma multiforme. Glioblastoma multiforme (GBM) is the most frequent brain tumor in adults. This lethal cancer is a challenge in neuro-oncology since patients almost invariably succumb to the disease. Intensive molecular studies have revealed a variety of deregulat...

Kasatori, N., 2000: A luminescent assay for natural cytotoxicity using K562 cells transfected with the plasmid producing alkaline phosphatase. A novel non-radioactive assay for natural cytotoxicity is described. A new target cell line was established by the transfection of K562 cells with a plasmid encoding DNA for secretion type of placental alkaline phosphatase(ALP). The assay was base...

Sinton, J.A., 1921: Relapsing Fever at Meshed, North-East Persia. Indian Medical Gazette 56(7): 241-250

Dweik, R.A.; Mehta, A.C.; Meeker, D.P.; Arroliga, A.C., 1996: Analysis of the safety of bronchoscopy after recent acute myocardial infarction. Acute myocardial infarction (AMI) is generally considered to increase the risk of flexible fiberoptic bronchoscopy (FFB). Currently, to our knowledge, no data in the literature support or challenge this concept. We conducted a retrospective chart...