The Mac
@TheMac
10 June, 08:37
An elegant way to precisely target 👉🏻optogenetic probes👈🏻👉🏻 to individual neurons is to use single-cell 👉🏻electroporation42,44 (Fig. 3). This involves using 👉🏻2-photon microscopy👈🏻 to target a plasmid-filled patch pipette to 👉🏻individual neurons in vivo and using 👉🏻electrical pulses33,34 to deliver the 👉🏻plasmid to the cell 👈🏻👉🏻under visual control.👉🏻 Neurons can be targeted this way based on their somatodendritic morphology👉🏻 (using ‘shadowimaging’33), 👉🏻their genetic identity👉🏻 (using GFP expression as a marker) 👉🏻or their functional properties (such as tuned responses to sensory stimuli) for subsequent 👉🏻optogenetic activation34.👈🏻
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10 June, 08:44
In response The Mac to his Publication
Yup.
People are now row-bots.
People are now row-bots.
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The Mac
@TheMac
10 June, 09:28
In response Photon 333 to her Publication
Hey Photon,
👉🏻momentarily👈🏻
👉🏻momentarily👈🏻
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10 June, 09:39
In response The Mac to his Publication
And, of course, that makes uninformed abuse okay.🙄
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The Mac
@TheMac
11 June, 12:06
In response Photon 333 to her Publication
2-photon microscopy
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The Mac
@TheMac
11 June, 12:12
In response The Mac to his Publication
Two-photon excitation microscopy (TPEF or 2PEF) is a fluorescence imaging technique that allows imaging of living tissue up to about one millimeter in thickness. Unlike traditional fluorescence microscopy, in which the excitation wavelength is shorter than the emission wavelength, two-photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light.
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The Mac
@TheMac
11 June, 12:13
In response The Mac to his Publication
A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent chemical compound that can re-emit light upon light excitation. Fluorophores typically contain several combined aromatic groups, or planar or cyclic molecules with several π bonds.
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Fluorophores are sometimes used alone, as a tracer in fluids, as a dye for staining of certain structures, as a substrate of enzymes, or as a probe or indicator (when its fluorescence is affected by environmental aspects such as polarity or ions). More generally they are covalently bonded to a macromolecule, serving as a marker (or dye, or tag, or reporter) for affine or bioactive reagents (antibodies, peptides, nucleic acids). Fluorophores are notably used to stain tissues, cells, or materials in a variety of analytical methods, i.e., fluorescent imaging and spectroscopy.
12:14 AM - Jun 11, 2021
In response The Mac to his Publication
Only people mentioned by TheMac in this post can reply
The Mac
@TheMac
11 June, 12:16
In response The Mac to his Publication
Fluorescein, via its amine-reactive isothiocyanate derivative fluorescein isothiocyanate (FITC), has been one of the most popular fluorophores. From antibody labeling, the applications have spread to nucleic acids thanks to carboxyfluorescein (FAM), TET, ...). Other historically common fluorophores are derivatives of rhodamine (TRITC), coumarin, and cyanine.[2] Newer generations of fluorophores, many of which are proprietary, often perform better, being more photostable, brighter, and/or less pH-sensitive than traditional dyes with comparable excitation and emission
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The Mac
@TheMac
11 June, 12:17
In response The Mac to his Publication
The fluorophore absorbs light energy of a specific wavelength and re-emits light at a longer wavelength. The absorbed wavelengths, energy transfer efficiency, and time before emission depend on both the fluorophore structure and its chemical environment, as the molecule in its excited state interacts with surrounding molecules. Wavelengths of maximum absorption (≈ excitation) and emission (for example, Absorption/Emission = 485 nm/517 nm) are the typical terms used to refer to a given fluorophore, but the whole spectrum may be important to consider.
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