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Apotome axiovert 200

Manufactured by Zeiss
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The Apotome Axiovert 200 is a laboratory instrument designed for optical sectioning microscopy. It utilizes a structured illumination technique to improve image contrast and optical sectioning capabilities. The core function of the Apotome Axiovert 200 is to enable high-resolution, three-dimensional imaging of fluorescently labeled samples.

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Lab products found in correlation

Ab169276, by Abcam (2 mentions) Mousemonoclonal 6 11 b1, by Merck Group (2 mentions) Hhpa036792, by Atlas Antibodies (2 mentions) Hpa 036791, by Atlas Antibodies (2 mentions) Lsm 880, by Zeiss (2 mentions) Alexa fluor 546, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Ab27043, by Abcam (1 mentions)

4 protocols using apotome axiovert 200

1

High-resolution Imaging of Ciliary Proteins

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High-resolution IFM was performed at the University Hospital of Muenster, Germany, following the laboratory protocols. The following antibodies were used: anti-DNAH5, anti-RSPH4A, anti-RSPH9, anti-CCDC39, anti-GAS8 and anti-DNAH116 . High-resolution immunofluorescence images were taken using a Zeiss Apotome Axiovert 200 (processed with AxioVision 4.8) or Zeiss LSM880 (processed with ZEN2 software) (see the Supplementary File).
Olm M.A., Marson F.A., Athanazio R.A., Nakagawa N.K., Macchione M., Loges N.T., Omran H., Rached S.Z., Bertuzzo C.S., Stelmach R., Saldiva P.H., Ribeiro J.D., Jones M.H, & Mauad T. (2019). Severe pulmonary disease in an adult primary ciliary dyskinesia population in Brazil. Scientific Reports, 9, 8693.
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2

Immunofluorescence Analysis of Respiratory Epithelial Cells

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Reciliated respiratory epithelial cells were suspended in cell culture medium and were blast by homogenizer for 3s to break the spheroids. Samples were spread onto glass slides, air-dried and stored at −80°C until use. Cells were treated with 4% paraformaldehyde, 0.1% Triton-X 100 and 1% BSA (all percentages are v/v) before incubation with primary (at least 1 h at room temperature (18–20°C) or overnight at 4°C) and secondary (30 min at room temperature) antibodies. Polyclonal anti-DNALI1, anti-DNAH5 and monoclonal mouse anti-acetylated-α-tubulin and monoclonal mouse gamma–tubulin antibodies were purchased from Sigma. Highly cross-absorbed secondary antibodies (Alexa Fluor 488, Alexa Fluor 546, Alex Fluor 633) were from Molecular Probes (Invitrogen). DNA was stained with DAPI (Sigma). Immunofluorescence images were taken with a Zeiss Apotome Axiovert 200 and processed with Openlab v5.1.
Li Y., Yagi H., Onuoha E.O., Damerla R.R., Francis R., Furutani Y., Tariq M., King S.M., Hendricks G., Cui C., Saydmohammed M., Lee D.M., Zahid M., Sami I., Leatherbury L., Pazour G.J., Ware S.M., Nakanishi T., Goldmuntz E., Tsang M, & Lo C.W. (2016). DNAH6 and Its Interactions with PCD Genes in Heterotaxy and Primary Ciliary Dyskinesia. PLoS Genetics, 12(2), e1005821.
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3

Immunofluorescence Analysis of Cell Lines

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Immunofluorescence and cell culture analyses were performed as
previously described.(Schmidts et al.,
2015) Briefly, IMCD3, hTERT-RPE, SH-SY5Y cells and patient as well as
control fibroblasts were cultured in DMEM-F12 medium with 5% FBS under standard
conditions. For ciliation experiments, cells were serum starved for 24 hours.
Primary Antibodies used: Golgi marker GM130 (mouse, ab169276, Abcam, USA), Golgi
marker 58K (ab27043 mouse, Abcam, USA), early endosome marker EEA1 (1G11
ab70521, mouse monoclonal, Abcam), PPP1R21 antibody (rabbit, HHPA036792 (ab1)
and HPA 036791 (ab2), Atlas antibodies, Sweden), acetylated tubulin (mouse
monoclonal 6-11-B1, Sigma, USA). In brief, cells were grown on coverslips,
washed with PBS and fixed 5 minutes in 5% PFA. After 3 washes, cells were
permeabilized using 0.1% Triton for 3 minutes, washed 3 times and the incubated
in 5% BSA for 1 hour at RT to bock unspecific reactions. Incubation in primary
antibodies was then performed for 1 hour (1:200 in 5% BSA), after 5 washes,
cells were incubated in corresponding fluorescence tagged secondary 1:1000
antibody dilutions for 30 minutes, washed 5 times in PBS and then mounted in
Vectashield with DAPI. mages were taken with a Zeiss Apotome Axiovert 200 and
processed with AxioVision 4.8 and Fiji software.
Rehman A.U., Najafi M., Kambouris M., Al‐Gazali L., Makrythanasis P., Rad A., Maroofian R., Rajab A., Stark Z., Hunter J.V., Bakey Z., Tokita M.J., He W., Vetrini F., Petersen A., Santoni F.A., Hamamy H., Wu K., Al‐Jasmi F., Helmstädter M., Arnold S.J., Xia F., Richmond C., Liu P., Karimiani E.G., Karami Madani G., Lunke S., El‐Shanti H., Eng C.M., Antonarakis S.E., Hertecant J., Walkiewicz M., Yang Y, & Schmidts M. (2018). Biallelic loss of function variants in PPP1R21 cause a neurodevelopmental syndrome with impaired endocytic function. Human Mutation, 40(3), 267-280.
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4

Immunofluorescence and Cell Culture Analysis

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Immunofluorescence and cell culture analyses were performed as previously described (Schmidts et al., 2015). Briefly, IMCD3, hTERT‐RPE, SH‐SY5Y cells and patient as well as control fibroblasts were cultured in DMEM‐F12 medium with 5% FBS under standard conditions. For ciliation experiments, cells were serum starved for 24 hr. Primary antibodies used: Golgi marker GM130 (mouse, ab169276, Abcam, USA), Golgi marker 58K (ab27043 mouse, Abcam, USA), early endosome marker EEA1 (1G11 ab70521, mouse monoclonal, Abcam), PPP1R21 antibody (rabbit, HHPA036792 (ab1) and HPA 036791 (ab2), Atlas antibodies, Sweden), acetylated tubulin (mouse monoclonal 6‐11‐B1, Sigma, USA). In brief, cells were grown on coverslips, washed with PBS and fixed 5 min in 5% PFA. After three washes, cells were permeabilized using 0.1% Triton for 3 min, washed three times and incubated in 5% BSA for 1 hr at RT to bock unspecific reactions. Incubation in primary antibodies was then performed for 1 hr (1:200 in 5% BSA), after five washes, cells were incubated in corresponding fluorescence tagged secondary 1:1,000 antibody dilutions for 30 min, washed five times in PBS and then mounted in Vectashield with DAPI. Images were taken with a Zeiss Apotome Axiovert 200 and processed with AxioVision 4.8 and Fiji software.
Rehman A.U., Najafi M., Kambouris M., Al‐Gazali L., Makrythanasis P., Rad A., Maroofian R., Rajab A., Stark Z., Hunter J.V., Bakey Z., Tokita M.J., He W., Vetrini F., Petersen A., Santoni F.A., Hamamy H., Wu K., Al‐Jasmi F., Helmstädter M., Arnold S.J., Xia F., Richmond C., Liu P., Karimiani E.G., Karami Madani G., Lunke S., El‐Shanti H., Eng C.M., Antonarakis S.E., Hertecant J., Walkiewicz M., Yang Y, & Schmidts M. (2018). Biallelic loss of function variants in PPP1R21 cause a neurodevelopmental syndrome with impaired endocytic function. Human Mutation, 40(3), 267-280.
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