Morphological changes accompanying cell lysis upon the thermal induction of prophages in lysogens were monitored by the time-lapse microscopy of living cells. To immobilize cells in one focal plane, samples (10 µL) withdrawn from the cultures of induced lysogens before the expected lysis time were placed on a microscopic slide on the upper surface of an agarose microslab (0.7% agarose in physiological saline) and covered with a coverslip. The microslabs were prepared as described previously [78 (link)]. Cells were immediately imaged using a Zeiss Axio M2 microscope with an oil immersion objective (100×, numerical aperture = 1.4). A time-lapse video was captured at five frames per second. The video was edited and scaled using the AxioVision 4 software package (Zeiss, Oberkochen, Germany). All images were stored as .zvi files and converted to TIFF or JPEG format as necessary.
Axio m2 microscope
Manufactured by Zeiss
Sourced in Germany
The Axio M2 microscope is a high-performance optical microscope designed for a wide range of applications. It features a modular design that allows for customization and integration with various accessories. The Axio M2 provides high-quality imaging and precise control over magnification, illumination, and other parameters.
Automatically generated - may contain errors
Lab products found in correlation
X citeseries 120 q mercury lamp, by Excelitas (2 mentions)
Axiovision 4, by Zeiss (1 mentions)
Toluidine blue o, by Merck Group (1 mentions)
Mab318, by Merck Group (1 mentions)
Vt1200s vibratome, by Leica (1 mentions)
A11001, by Thermo Fisher Scientific (1 mentions)
Ultraflextreme mass spectrometer, by Bruker (1 mentions)
Zen software, by Zeiss (1 mentions)
Ab cam icc5 camera, by Excelitas (1 mentions)
Hal 100 halogen illuminator, by Zeiss (1 mentions)
Zen software version 2 blue edition, by Zeiss (1 mentions)
6 protocols using axio m2 microscope
1
Time-lapse Microscopy of Prophage Induction
2
Histological Analysis of VGAT-Cre Mouse Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animals were perfused using 4% paraformaldehyde (PFA) in phosphate-buffered saline (PBS) and brains were extracted and fixed in PFA-PBST. A vibratome was used to collect 60 micron coronal slices stored in PBS. Images show DAPI staining in blue and EYFP in green. 2x and 10x images were taken on a Zeiss Axio M2 microscope. In addition to the histology from mice in which we performed electrophysiological recordings (Figure 1—figure supplement 1,2,5 ; Figure 4—figure supplement 2 ), we also injected an additional VGAT-Cre mouse with the same halorhodopsin virus and performed histology at higher resolution. The same histological methods were used, except that slices were 65 microns thick. These images were taken on a Zeiss 750 confocal microscope (Figure 4—figure supplement 3 ). Cell counting was performed manually, and the proportion of cells that were positive (i.e., were surrounded by a fluorescent ring) was reported with error bars indicating the 95% confidence intervals calculated theoretically from the binomial distribution.
3
Toluidine Blue O Staining of Root Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Toluidine blue O 0.1% was made by dissolving 0.1 g Toluidine blue O (Sigma, St Louis, MO, USA) in 100 ml pure water. The vacuum-dried cross-sections of roots were attached to the glass slide and transferred to a 10-mm petri dish. The tissue sections were immersed in the staining solution and incubated for 5 min at room temperature. Staining solution was gently removed using a pipette, and the tissue sections were rinsed with pure water to remove excess stain. The stained sections were mounted in pure water under a coverslip and optical images of tissue sections were acquired with an Axio M2 microscope (Zeiss, Oberkochen, Germany).
4
Validating DREADD Expression in Dopamine Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After all experiments were completed, viral expression and localization was verified via histologic analysis. Dat-cre animals were perfused with phosphate buffered saline followed by neutral buffered formalin. The brains were postfixed in formalin overnight, and sliced at 60 μm using a Leica VT1200 S vibratome (Leica Microsystems Inc.). Specific expression of DREADDs in DA neurons was confirmed by colocalization of mCherry (from AAV expression) with immunohistochemical staining for tyrosine hydroxylase (TH), a marker of DA neurons (mouse anti-TH, 1:1000 dilution, Millipore catalog #MAB318), using the secondary antibody of goat anti-mouse conjugated to Alexa Fluor 488 (1:200 dilution, catalog #A-11001, Invitrogen). Cells were counterstained with 4’,6-diamidino-2-phenylindole (DAPI; catalog #H-1200, Vectashield) for nuclear visualization. Images were taken with a Zeiss Axio M2 microscope (Zeiss). Confirmation of viral expression in the correct brain region was performed by comparing images to a Mouse Brain Atlas (Paxinos et al., 2001 ).
5
Microscopy-Guided Mass Spectrometry Imaging
6
Brightfield and Fluorescence Imaging
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!