Log In Sign up
The largest database of trusted experimental protocols

Apotome attachment

Manufactured by Zeiss
Visit Supplier
Sourced in Germany

The ApoTome attachment is a specialized optical system designed for Carl Zeiss microscopes. It uses a structured illumination technique to improve the contrast and optical sectioning of samples, allowing for the acquisition of high-quality, optically sectioned images.

Automatically generated - may contain errors

Lab products found in correlation

Axio imager z1 microscope, by Zeiss (3 mentions) Axio imager z2 microscope, by Zeiss (2 mentions) Nis element, by Nikon (2 mentions) Zen 3, by Zeiss (2 mentions) Eclipse ni2, by Nikon (2 mentions) Dako mounting media, by Agilent Technologies (1 mentions) Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions) Axiovision software, by Zeiss (1 mentions) Axiocam mr, by Zeiss (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Tritc labeled phalloidin, by Merck Group (1 mentions) Rabbit anti dcp 1 asp216, by Cell Signaling Technology (1 mentions) Zen software, by Zeiss (1 mentions) Orca r2 camera, by Hamamatsu Photonics (1 mentions) Axio imager m2 microscope, by Zeiss (1 mentions) Axio observer z1 microscope, by Zeiss (1 mentions) Stereo discovery v12 microscope, by Zeiss (1 mentions)

Close spelling variants of this product

Apotome (432 citations) Apotome.2 attachment (2 citations)

10 protocols using apotome attachment

1

Epifluorescent Microscopy with ApoTome

Check if the same lab product or an alternative is used in the 5 most similar protocols
Representative images were taken using an epifluorescent Axio Imager Z1 microscope with ApoTome attachment (Carl Zeiss, Oberkochen, Germany).
Meehan G.R., McGonigal R., Cunningham M.E., Wang Y., Barrie J.A., Halstead S.K., Gourlay D., Yao D, & Willison H.J. (2018). Differential binding patterns of anti-sulfatide antibodies to glial membranes. Journal of Neuroimmunology, 323, 28-35.
+ Open protocol
+ Expand
2

Immunofluorescence Microscopy of Transfected Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated in 6-well plates and grown for 24 h. They were then transfected and 6 h after transfection they were transferred (diluted 1:10) to coverslips in a 24-well plate coated with poly-D-lysine (PDL) and collagen. Cells then grew for 24–72 h before fixation with 4% PFA/PBS for 20 min at RT depending on the experiment. Permeabilization of the cell membrane was done with 1% Triton-X-100/PBS for 5 min at RT. Blocking was done with 10% Normal Goat Serum in PBS, for 1 h at RT. The primary antibody incubations were done at the already mentioned concentrations, in 1%BSA/PBS for 2 h at RT. The secondary antibody incubation was done in 1% BSA/PBS for 1 h. Mounting was done using 40 µl of Dako mounting media per slide, following a 4° incubation overnight. Immunofluorescence images were acquired using a Zeiss fluorescence microscope with Apotome attachment (Axio imager z1 stand).
Garcia Morato J., Hans F., von Zweydorf F., Feederle R., Elsässer S.J., Skodras A.A., Gloeckner C.J., Buratti E., Neumann M, & Kahle P.J. (2022). Sirtuin-1 sensitive lysine-136 acetylation drives phase separation and pathological aggregation of TDP-43. Nature Communications, 13, 1223.
+ Open protocol
+ Expand
3

Quantitative Fluorescence Microscopy Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Images of immunofluorescence were captured using an Axio-imager Z2 microscope fitted with an apotome attachment for structured illumination (Carl Zeiss MicroImaging) and a Nikon Eclipse Ni2. 20X (0.8 NA), 40X (0.95 NA), and 63X (1.4 NA) objectives were used. Images were taken using Zen 3.2 (Zeiss) or NIS-Elements (Nikon). For measurements of AIS streptavidin fluorescence intensity, 20 neurons per timepoint per replicate were imaged and line scans were drawn using Zen 3.2 software. AIS were identified by immunostaining using known AIS proteins (AnkG, β4 spectrin, Nfasc, or NrCAM). For the analysis of cerebellar pinceau, a region of interest including the Purkinje neuron AIS (labeled by Nfasc) was manually drawn. The fluorescence intensity for Kv1.2 was measured for each region of interest and normalized to the area. All measurements were taken with the same exposure times and immunolabeling was also performed at the same time. Images were exported to Fiji, Adobe Photoshop, and Adobe illustrator for figure presentation. Some figures were generated using Biorender.
Ogawa Y., Lim B.C., George S., Oses-Prieto J.A., Rasband J.M., Eshed-Eisenbach Y., Hamdan H., Nair S., Boato F., Peles E., Burlingame A.L., Van Aelst L, & Rasband M.N. (2023). Antibody-directed extracellular proximity biotinylation reveals that Contactin-1 regulates axo-axonic innervation of axon initial segments. Nature Communications, 14, 6797.
+ Open protocol
+ Expand
4

Microscopy Techniques for Germ Cell Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Experimental procedures for electron and fluorescence microscopy were performed as described earlier (Pertceva et al., 2010 (link)). The primary antibodies were: monoclonal rabbit anti-VASA antibodies from Santa Cruz Biotechnology (1:300), rabbit anti-Dcp-1 (Asp216) from Cell Signaling Technology. The secondary antibodies were goat anti-rabbit conjugated to AlexaFluor-488 (1:500; Invitrogen #A-11001). TRITC-labeled phalloidin (Sigma-Aldrich #P1951) was used at 1:100 dilution to visualize F-actin as described previously (Guild et al., 1997 (link)). The LysoTracker assay was performed as described in Dorogova et al. (2014) (link) (LysoTracker red DND-99 (Invitrogen, Molecular Probes, Basel, Switzerland). Ovaries were embedded with ProLong Gold anti-fade reagent with DAPI. Images were obtained using an AxioImager Z1 microscope with ApoTome attachment (Zeiss), AxioCam MR and AxioVision software (Zeiss, Germany).
Dorogova N.V., Galimova Y.A., Bolobolova E.U., Baricheva E.M, & Fedorova S.A. (2020). Loss of Drosophila E3 Ubiquitin Ligase Hyd Promotes Extra Mitosis in Germline Cysts and Massive Cell Death During Oogenesis. Frontiers in Cell and Developmental Biology, 8, 600868.
+ Open protocol
+ Expand
5

Quantitative Nerve Terminal Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols
All imaging was performed using a Zeiss Axio Z1 Imager with an Apotome attachment (removed for imaging sections, attached for imaging whole-mount tissue) and captured with Zen Blue Edition software (version 6.1.7). Each stain was performed in duplicate for each mouse and all tissue was coded prior to imaging. Every distal nerve identified by MBP and BTx staining was imaged in each diaphragm/TS. Z-stacks with a 0.4 μm interval were taken using a 63 × oil objective. For these images, 24–89 nerve terminals and 5–44 distal nerves were analysed per mouse. The distal nerve was defined as the nerve terminal, first internode and first NoR identified by a gap in MBP staining. When assessing complement deposition, if deposits were detected overlying the distal internode and/or NoR, this was scored as positive. To assess the integrity of proteins at the NoR, presence/absence analysis was performed. When assessing the integrity of paranodal markers AnkB, CASPR1 and NF155, presence was defined as both domains and absence was defined as a single domain or complete loss of staining.
Campbell C.I., McGonigal R., Barrie J.A., Delaere J., Bracke L., Cunningham M.E., Yao D., Delahaye T., Van de Walle I, & Willison H.J. (2022). Complement inhibition prevents glial nodal membrane injury in a GM1 antibody-mediated mouse model. Brain Communications, 4(6), fcac306.
+ Open protocol
+ Expand
6

Quantifying Retrograde Labeled and Neuropeptide-Expressing Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
The number of CTB+, AVP-ir, and CTB/AVP co-labeled neurons were counted using ImageJ (NIH; imagej.nih.gov/ij). Images were acquired using an Axiolmager fluorescence microscope with ApoTome attachment (Carl Zeiss Microscopy GmbH, Jena, Germany), Orca-R2 camera (Hamamatsu, Bridgewater, NJ, USA) and Zen software (Carl Zeiss Microscopy GmbH, Jena, Germany). Tiled confocal fluorescent images (2 × 2mm; 20μm Z-plane) encompassing the posterior bed nucleus of the stria terminalis (pBNST) and posterodorsal medial amygdala (MePD) were Z-stacked, and separated into red, green, and yellow channels using the ImageJ split channel function. CTB+ neurons were counted using the cell counter function in ImageJ from the red channel, AVP-ir neurons were counted from the green channel, and colabeled neurons were quantified from the yellow channel. The total number of colabeled cells in each image was then averaged across three non-overlapping sections in an evenly spaced series (90pm apart) for both the pBNST and MePD. Many AVP-ir cells were observed in the paraventricular (PVN) and supraoptic (SON) nuclei of the hypothalamus, however in no cases were any back-labeled CTB+ cell bodies co-expressing AVP observed in these regions. Thus, these regions were not further analyzed.
DiBenedictis B.T., Cheung H.K., Nussbaum E.R, & Veenema A.H. (2019). Involvement of ventral pallidal vasopressin in the sex-specific regulation of sociosexual motivation in rats. Psychoneuroendocrinology, 111, 104462.
+ Open protocol
+ Expand
7

Quantifying Microglia and Macrophages in Hippocampus

Check if the same lab product or an alternative is used in the 5 most similar protocols
The analysis of resident microglia and blood‐derived macrophages was performed by quantifying Cx3cr1‐GFP‐positive (Cx3cr1‐GFP+) and Ccr2‐RFP‐positive (Ccr2‐RFP+) cells in the entire dorsal DG region of the hippocampus (the granule cell layer including the subgranular zone) and the entire dorsal CA region of the hippocampus (the pyramidal cell layer from CA1‐3) (Fanselow and Dong, 2010; Qiu et al., 2007). For quantification of immunopositive cells in the DG and the CA regions, ten consecutive 0.5μm z‐stack images of the sections were acquired using Axio Imager M2 microscope with Apotome attachment (Carl Zeiss) at 20× magnification and compressed in a single image (Kadam et al., 2015). All immunopositive cells were counted in three to four sections per animal spaced 400 μm apart. The total number of cells was obtained by multiplying the number of cells by the sampling fractions. To evaluate the proliferation of microglia, 100 Cx3cr1‐GFP+ cells were quantified and examined for coexpression of Ki67.
Umekawa T., Osman A.M., Han W., Ikeda T, & Blomgren K. (2015). Resident microglia, rather than blood‐derived macrophages, contribute to the earlier and more pronounced inflammatory reaction in the immature compared with the adult hippocampus after hypoxia‐ischemia. Glia, 63(12), 2220-2230.
+ Open protocol
+ Expand
8

Quantitative Cell Invasion Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
For cell invasion assays, cells were cultured in the medium used for tumorsphere formation supplemented with 0.2% methylcellulose in a non-adhesive convex environment for 12 h at 37 °C and 5% CO2. Tumorspheres were mixed with collagen matrix (2.5 mg/ml) and incubated for 30 min at 37 °C prior to microscopic analysis. Time-lapse microscopy imaging was performed on a Zeiss AxioObserver Z1 microscope with a Plan-Apochromat ×40 /1.3 (NA = 1.3, working distance = 0.21 mm), a camera, and an Apotome attachment (Carl Zeiss, Germany). Mosaic images were collected using AxioVision software over a period of 20 h with a time resolution of 30 min.
Herreros-Pomares A., de-Maya-Girones J.D., Calabuig-Fariñas S., Lucas R., Martínez A., Pardo-Sánchez J.M., Alonso S., Blasco A., Guijarro R., Martorell M., Escorihuela E., Chiara M.D., Duréndez E., Gandía C., Forteza J., Sirera R., Jantus-Lewintre E., Farràs R, & Camps C. (2019). Lung tumorspheres reveal cancer stem cell-like properties and a score with prognostic impact in resected non-small-cell lung cancer. Cell Death & Disease, 10(9), 660.
+ Open protocol
+ Expand
9

Brightfield Imaging of Live Zebrafish Embryos

Check if the same lab product or an alternative is used in the 5 most similar protocols
Brightfield images of live embryos were taken on the Zeiss SteREO Discovery V12 microscope at ×25 or the Zeiss Axio Imager Z1 microscope with the ×5 objective. Live embryos were anesthetized using a non-lethal dose of MS-222. Fixed embryos were deyolked in PBS, side-mounted in 80 % glycerol/20 % PBS, and imaged with the ×20 objective of a Zeiss Axio Imager Z1 microscope with a Zeiss ApoTome attachment and optimized by averaging five frames. For images where fluorescence levels were to be compared, exposure times were kept constant throughout the imaging of that experiment. All image modifications were performed in Adobe Photoshop with Gaussian filtering (0.3 pixel) and unsharp mask (50 %, 1 pixel) prior to being collated in Adobe Illustrator.
Jenkins M.H., Alrowaished S.S., Goody M.F., Crawford B.D, & Henry C.A. (2016). Laminin and Matrix metalloproteinase 11 regulate Fibronectin levels in the zebrafish myotendinous junction. Skeletal Muscle, 6, 18.
+ Open protocol
+ Expand
10

Immunofluorescence Microscopy Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Images of immunofluorescence were captured using an Axio-imager Z2 microscope fitted with an apotome attachment for structured illumination (Carl Zeiss MicroImaging) and a Nikon Eclipse Ni2. 20X (0.8 NA), 40X (0.95 NA), and 63X (1.4 NA) objectives were used. Images were taken using Zen 3.2 (Zeiss) or NIS-Elements (Nikon). For measurements of AIS streptavidin fluorescence intensity, 20 neurons per timepoint per replicate were imaged and line scans were drawn using Zen 3.2 software. Images were exported to Fiji, Adobe Photoshop, and Adobe illustrator for figure presentation. Some figures were generated using Biorender.
Ogawa Y., Lim B.C., George S., Oses-Prieto J.A., Rasband J.M., Eshed-Eisenbach Y., Nair S., Boato F., Peles E., Burlingame A.L., Van Aelst L, & Rasband M.N. (2023). Antibody-directed extracellular proximity biotinylation reveals Contactin-1 regulates axo-axonic innervation of axon initial segments. bioRxiv.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!