MITOCHINDRIA
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All We have isolated mitochondria from human cells and want to see them in confocal microscope. Can someone tell which objective should i use for this Regards |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Could you please tell what exactly you are going to image and what is the label? Are you going to perform some quantitative imgaing or just make sure that your sample really contains mitochondria? What is the cell type? Mitchondria are well visible for 1.2 NA objective (TMRE staining, lymphocytes) and qualitatively visible while using NA 1 objective if cells are spreaded (MitoTracker Green, Vero cells) Best Sergey Tauger PhD student Cell motility lab. Dept. of Biology Moscow State University |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I am also trying to localize mitochondria in fish macrophages using mitotracker. What I am getting is greenish tinge of mitotracker at x 100 objective. Can u please suggest me if there is some software for digital zooming Regards Chaitali On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Could you please tell what exactly you are going to image and what is the > label? > Are you going to perform some quantitative imgaing or just make sure that > your > sample really contains mitochondria? What is the cell type? > > Mitchondria are well visible for 1.2 NA objective (TMRE staining, > lymphocytes) and > qualitatively visible while using NA 1 objective if cells are spreaded > (MitoTracker > Green, Vero cells) > > Best > Sergey Tauger > PhD student > Cell motility lab. > Dept. of Biology > Moscow State University > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear even i am trying to but getting nothing On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < [hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I am also trying to localize mitochondria in fish macrophages using > mitotracker. What I am getting is greenish tinge of mitotracker at x 100 > objective. Can u please suggest me if there is some software for digital > zooming > Regards > Chaitali > > > On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Could you please tell what exactly you are going to image and what is the > > label? > > Are you going to perform some quantitative imgaing or just make sure that > > your > > sample really contains mitochondria? What is the cell type? > > > > Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > lymphocytes) and > > qualitatively visible while using NA 1 objective if cells are spreaded > > (MitoTracker > > Green, Vero cells) > > > > Best > > Sergey Tauger > > PhD student > > Cell motility lab. > > Dept. of Biology > > Moscow State University > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Anyone please help On Mon, Feb 24, 2014 at 4:36 PM, Amandeep Walia <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear even i am trying to but getting nothing > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > [hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > I am also trying to localize mitochondria in fish macrophages using > > mitotracker. What I am getting is greenish tinge of mitotracker at x 100 > > objective. Can u please suggest me if there is some software for digital > > zooming > > Regards > > Chaitali > > > > > > On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> > wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Could you please tell what exactly you are going to image and what is > the > > > label? > > > Are you going to perform some quantitative imgaing or just make sure > that > > > your > > > sample really contains mitochondria? What is the cell type? > > > > > > Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > > lymphocytes) and > > > qualitatively visible while using NA 1 objective if cells are spreaded > > > (MitoTracker > > > Green, Vero cells) > > > > > > Best > > > Sergey Tauger > > > PhD student > > > Cell motility lab. > > > Dept. of Biology > > > Moscow State University > > > > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** You don't need a specific objective to see mitochondria. You first need a good staining. Best results are achieved with mitochondrial targeted FPs or through immunofluorescence of mitochondrial proteins (TOM20, TOM40, HSP60 and many others). Otherwise, if you want to use mitotracker dyes, you need to set up dye concentration and loading time to obtain good results. Diego De Stefani, PhD Department of Biomedical Sciences University of Padova Il 24/feb/2014 13:53 "Chaitali Banerjee" <[hidden email]> ha scritto: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Anyone please help > > > On Mon, Feb 24, 2014 at 4:36 PM, Amandeep Walia <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Dear even i am trying to but getting nothing > > > > > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > > [hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > I am also trying to localize mitochondria in fish macrophages using > > > mitotracker. What I am getting is greenish tinge of mitotracker at x > 100 > > > objective. Can u please suggest me if there is some software for > digital > > > zooming > > > Regards > > > Chaitali > > > > > > > > > On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> > > wrote: > > > > > > > ***** > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > ***** > > > > > > > > Could you please tell what exactly you are going to image and what is > > the > > > > label? > > > > Are you going to perform some quantitative imgaing or just make sure > > that > > > > your > > > > sample really contains mitochondria? What is the cell type? > > > > > > > > Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > > > lymphocytes) and > > > > qualitatively visible while using NA 1 objective if cells are > spreaded > > > > (MitoTracker > > > > Green, Vero cells) > > > > > > > > Best > > > > Sergey Tauger > > > > PhD student > > > > Cell motility lab. > > > > Dept. of Biology > > > > Moscow State University > > > > > > > > > > |
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In reply to this post by Amandeep Walia-2
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Given the gmail accounts and lack of affiliation, are you and Chaitali working together? Are you looking for signal or resolution? Do you have a suitable camera? An overly corrected objective, low NA or phase objectives will all reduce signal. Mitotrackers in zebrafish can be detected in widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and easily recorded with NA 1.0-1.2. I hope you are not trying to look through a plastic culture dish. Do you have any experience with live imaging, microscopy or dye loading? Have you looked in the literature? There are many papers using Mitotrackers, and in zebrafish. Providing a description of your instrumentation, experimental intent and sample preparations when you ask for assistance will avoid a game of “20 Questions”. Mitotrackers require testing for concentration, incubation times, temperature, etc. before embarking on any experiment but the literature will provide starting points. Overloading can reduce intensity (by disrupting mitochondrial function), and is a common beginner’s mistake. try 25 nM to 100 nM, 20-30 min. or shorter. MT Red is brighter, use no more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit voltage sensitive. Digital zoom will not improve signal, it sometimes helps with visibility on the screen. Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Cellular Morphology Core Center on Human Development and Disability Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 [hidden email] On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear even i am trying to but getting nothing > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > [hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> I am also trying to localize mitochondria in fish macrophages using >> mitotracker. What I am getting is greenish tinge of mitotracker at x 100 >> objective. Can u please suggest me if there is some software for digital >> zooming >> Regards >> Chaitali >> >> >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Could you please tell what exactly you are going to image and what is the >>> label? >>> Are you going to perform some quantitative imgaing or just make sure that >>> your >>> sample really contains mitochondria? What is the cell type? >>> >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining, >>> lymphocytes) and >>> qualitatively visible while using NA 1 objective if cells are spreaded >>> (MitoTracker >>> Green, Vero cells) >>> >>> Best >>> Sergey Tauger >>> PhD student >>> Cell motility lab. >>> Dept. of Biology >>> Moscow State University >>> >> |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I am a PhD stuent from University of Dlehi, India. Currently I am performing some experiments with mito tracker and ER tracker and was studying mitochondrial movement towards ER. When i tried co localization i was nt happy with my results. I am using 10 nm of mitotracker & er tracker.When i looked into literature i got sm pictures like what i hv got, but I found that they further digitally magnified the images several fold. I tried with the photoshop & imagej to get the best magnificatn. But still nt much convincd. Any suggestn please. Regards Chaitali Banerjee PhD student Department of Zoology University of Delhi India On Mon, Feb 24, 2014 at 8:45 PM, Glen MacDonald <[hidden email]>wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Given the gmail accounts and lack of affiliation, are you and Chaitali > working together? Are you looking for signal or resolution? Do you have a > suitable camera? An overly corrected objective, low NA or phase objectives > will all reduce signal. Mitotrackers in zebrafish can be detected in > widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and > easily recorded with NA 1.0-1.2. I hope you are not trying to look through > a plastic culture dish. Do you have any experience with live imaging, > microscopy or dye loading? Have you looked in the literature? There are > many papers using Mitotrackers, and in zebrafish. > > Providing a description of your instrumentation, experimental intent and > sample preparations when you ask for assistance will avoid a game of "20 > Questions". > > Mitotrackers require testing for concentration, incubation times, > temperature, etc. before embarking on any experiment but the literature > will provide starting points. Overloading can reduce intensity (by > disrupting mitochondrial function), and is a common beginner's mistake. > try 25 nM to 100 nM, 20-30 min. or shorter. MT Red is brighter, use no > more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit voltage > sensitive. > > Digital zoom will not improve signal, it sometimes helps with visibility > on the screen. > > Glen MacDonald > Core for Communication Research > Virginia Merrill Bloedel Hearing Research Center > Cellular Morphology Core > Center on Human Development and Disability > Box 357923 > University of Washington > Seattle, WA 98195-7923 USA > (206) 616-4156 > [hidden email] > > > > > > > > On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Dear even i am trying to but getting nothing > > > > > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > > [hidden email]> wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** > >> > >> I am also trying to localize mitochondria in fish macrophages using > >> mitotracker. What I am getting is greenish tinge of mitotracker at x 100 > >> objective. Can u please suggest me if there is some software for digital > >> zooming > >> Regards > >> Chaitali > >> > >> > >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> > wrote: > >> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** > >>> > >>> Could you please tell what exactly you are going to image and what is > the > >>> label? > >>> Are you going to perform some quantitative imgaing or just make sure > that > >>> your > >>> sample really contains mitochondria? What is the cell type? > >>> > >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining, > >>> lymphocytes) and > >>> qualitatively visible while using NA 1 objective if cells are spreaded > >>> (MitoTracker > >>> Green, Vero cells) > >>> > >>> Best > >>> Sergey Tauger > >>> PhD student > >>> Cell motility lab. > >>> Dept. of Biology > >>> Moscow State University > >>> > >> > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Chaitali, Did you perform the control sample along with your experimental sample, which microscope/ software you used for colocalization analysis? Rgds Manish Imaging Facility Manager IGIB, New Delhi On Tue, Feb 25, 2014 at 11:13 AM, Chaitali Banerjee < [hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I am a PhD stuent from University of Dlehi, India. Currently I am > performing some experiments with mito tracker and ER tracker and was > studying mitochondrial movement towards ER. When i tried co localization i > was nt happy with my results. I am using 10 nm of mitotracker & er > tracker.When > i looked into literature i got sm pictures like what i hv got, > but I found that they further digitally magnified the images several fold. > I tried with the photoshop & imagej to get the best magnificatn. But still > nt much convincd. Any suggestn please. > Regards > Chaitali Banerjee > PhD student > Department of Zoology > University of Delhi > India > > > On Mon, Feb 24, 2014 at 8:45 PM, Glen MacDonald <[hidden email] > >wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Given the gmail accounts and lack of affiliation, are you and Chaitali > > working together? Are you looking for signal or resolution? Do you > have a > > suitable camera? An overly corrected objective, low NA or phase > objectives > > will all reduce signal. Mitotrackers in zebrafish can be detected in > > widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and > > easily recorded with NA 1.0-1.2. I hope you are not trying to look > through > > a plastic culture dish. Do you have any experience with live imaging, > > microscopy or dye loading? Have you looked in the literature? There are > > many papers using Mitotrackers, and in zebrafish. > > > > Providing a description of your instrumentation, experimental intent and > > sample preparations when you ask for assistance will avoid a game of "20 > > Questions". > > > > Mitotrackers require testing for concentration, incubation times, > > temperature, etc. before embarking on any experiment but the literature > > will provide starting points. Overloading can reduce intensity (by > > disrupting mitochondrial function), and is a common beginner's mistake. > > try 25 nM to 100 nM, 20-30 min. or shorter. MT Red is brighter, use no > > more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit > voltage > > sensitive. > > > > Digital zoom will not improve signal, it sometimes helps with visibility > > on the screen. > > > > Glen MacDonald > > Core for Communication Research > > Virginia Merrill Bloedel Hearing Research Center > > Cellular Morphology Core > > Center on Human Development and Disability > > Box 357923 > > University of Washington > > Seattle, WA 98195-7923 USA > > (206) 616-4156 > > [hidden email] > > > > > > > > > > > > > > > > On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Dear even i am trying to but getting nothing > > > > > > > > > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > > > [hidden email]> wrote: > > > > > >> ***** > > >> To join, leave or search the confocal microscopy listserv, go to: > > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > >> ***** > > >> > > >> I am also trying to localize mitochondria in fish macrophages using > > >> mitotracker. What I am getting is greenish tinge of mitotracker at x > 100 > > >> objective. Can u please suggest me if there is some software for > digital > > >> zooming > > >> Regards > > >> Chaitali > > >> > > >> > > >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> > > wrote: > > >> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go to: > > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > >>> ***** > > >>> > > >>> Could you please tell what exactly you are going to image and what is > > the > > >>> label? > > >>> Are you going to perform some quantitative imgaing or just make sure > > that > > >>> your > > >>> sample really contains mitochondria? What is the cell type? > > >>> > > >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > >>> lymphocytes) and > > >>> qualitatively visible while using NA 1 objective if cells are > spreaded > > >>> (MitoTracker > > >>> Green, Vero cells) > > >>> > > >>> Best > > >>> Sergey Tauger > > >>> PhD student > > >>> Cell motility lab. > > >>> Dept. of Biology > > >>> Moscow State University > > >>> > > >> > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Manish I have used both control and treated sample. I am using Zeiss automated microscope and using 100x. Can u please tell me in which campus you are working-mall road or mathura road. I am in north campus,Delhi. I can come to meet u then. Regards Chaitali On Tue, Feb 25, 2014 at 3:14 PM, Manish Kumar <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi Chaitali, > Did you perform the control sample along with your experimental sample, > which microscope/ software you used for colocalization analysis? > > Rgds > Manish > Imaging Facility Manager > IGIB, New Delhi > > > On Tue, Feb 25, 2014 at 11:13 AM, Chaitali Banerjee < > [hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > I am a PhD stuent from University of Dlehi, India. Currently I am > > performing some experiments with mito tracker and ER tracker and was > > studying mitochondrial movement towards ER. When i tried co localization > i > > was nt happy with my results. I am using 10 nm of mitotracker & er > > tracker.When > > i looked into literature i got sm pictures like what i hv got, > > but I found that they further digitally magnified the images several > fold. > > I tried with the photoshop & imagej to get the best magnificatn. But > still > > nt much convincd. Any suggestn please. > > Regards > > Chaitali Banerjee > > PhD student > > Department of Zoology > > University of Delhi > > India > > > > > > On Mon, Feb 24, 2014 at 8:45 PM, Glen MacDonald < > [hidden email] > > >wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Given the gmail accounts and lack of affiliation, are you and Chaitali > > > working together? Are you looking for signal or resolution? Do you > > have a > > > suitable camera? An overly corrected objective, low NA or phase > > objectives > > > will all reduce signal. Mitotrackers in zebrafish can be detected in > > > widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and > > > easily recorded with NA 1.0-1.2. I hope you are not trying to look > > through > > > a plastic culture dish. Do you have any experience with live imaging, > > > microscopy or dye loading? Have you looked in the literature? There > are > > > many papers using Mitotrackers, and in zebrafish. > > > > > > Providing a description of your instrumentation, experimental intent > and > > > sample preparations when you ask for assistance will avoid a game of > "20 > > > Questions". > > > > > > Mitotrackers require testing for concentration, incubation times, > > > temperature, etc. before embarking on any experiment but the literature > > > will provide starting points. Overloading can reduce intensity (by > > > disrupting mitochondrial function), and is a common beginner's mistake. > > > try 25 nM to 100 nM, 20-30 min. or shorter. MT Red is brighter, use > no > > > more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit > > voltage > > > sensitive. > > > > > > Digital zoom will not improve signal, it sometimes helps with > visibility > > > on the screen. > > > > > > Glen MacDonald > > > Core for Communication Research > > > Virginia Merrill Bloedel Hearing Research Center > > > Cellular Morphology Core > > > Center on Human Development and Disability > > > Box 357923 > > > University of Washington > > > Seattle, WA 98195-7923 USA > > > (206) 616-4156 > > > [hidden email] > > > > > > > > > > > > > > > > > > > > > > > > On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[hidden email]> wrote: > > > > > > > ***** > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > ***** > > > > > > > > Dear even i am trying to but getting nothing > > > > > > > > > > > > > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > > > > [hidden email]> wrote: > > > > > > > >> ***** > > > >> To join, leave or search the confocal microscopy listserv, go to: > > > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > >> ***** > > > >> > > > >> I am also trying to localize mitochondria in fish macrophages using > > > >> mitotracker. What I am getting is greenish tinge of mitotracker at x > > 100 > > > >> objective. Can u please suggest me if there is some software for > > digital > > > >> zooming > > > >> Regards > > > >> Chaitali > > > >> > > > >> > > > >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email]> > > > wrote: > > > >> > > > >>> ***** > > > >>> To join, leave or search the confocal microscopy listserv, go to: > > > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > >>> ***** > > > >>> > > > >>> Could you please tell what exactly you are going to image and what > is > > > the > > > >>> label? > > > >>> Are you going to perform some quantitative imgaing or just make > sure > > > that > > > >>> your > > > >>> sample really contains mitochondria? What is the cell type? > > > >>> > > > >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > > >>> lymphocytes) and > > > >>> qualitatively visible while using NA 1 objective if cells are > > spreaded > > > >>> (MitoTracker > > > >>> Green, Vero cells) > > > >>> > > > >>> Best > > > >>> Sergey Tauger > > > >>> PhD student > > > >>> Cell motility lab. > > > >>> Dept. of Biology > > > >>> Moscow State University > > > >>> > > > >> > > > > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** you can meet me at Mathura road campus. On Tue, Feb 25, 2014 at 4:16 PM, Chaitali Banerjee < [hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi Manish > I have used both control and treated sample. I am using Zeiss automated > microscope and using 100x. Can u please tell me in which campus you are > working-mall road or mathura road. I am in north campus,Delhi. I can come > to meet u then. > Regards > Chaitali > > > On Tue, Feb 25, 2014 at 3:14 PM, Manish Kumar <[hidden email]> > wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi Chaitali, > > Did you perform the control sample along with your experimental sample, > > which microscope/ software you used for colocalization analysis? > > > > Rgds > > Manish > > Imaging Facility Manager > > IGIB, New Delhi > > > > > > On Tue, Feb 25, 2014 at 11:13 AM, Chaitali Banerjee < > > [hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > I am a PhD stuent from University of Dlehi, India. Currently I am > > > performing some experiments with mito tracker and ER tracker and was > > > studying mitochondrial movement towards ER. When i tried co > localization > > i > > > was nt happy with my results. I am using 10 nm of mitotracker & er > > > tracker.When > > > i looked into literature i got sm pictures like what i hv got, > > > but I found that they further digitally magnified the images several > > fold. > > > I tried with the photoshop & imagej to get the best magnificatn. But > > still > > > nt much convincd. Any suggestn please. > > > Regards > > > Chaitali Banerjee > > > PhD student > > > Department of Zoology > > > University of Delhi > > > India > > > > > > > > > On Mon, Feb 24, 2014 at 8:45 PM, Glen MacDonald < > > [hidden email] > > > >wrote: > > > > > > > ***** > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > ***** > > > > > > > > Given the gmail accounts and lack of affiliation, are you and > Chaitali > > > > working together? Are you looking for signal or resolution? Do you > > > have a > > > > suitable camera? An overly corrected objective, low NA or phase > > > objectives > > > > will all reduce signal. Mitotrackers in zebrafish can be detected in > > > > widefield or confocal with a 40X/.75 (or 20X + 2X zoom) air lens, and > > > > easily recorded with NA 1.0-1.2. I hope you are not trying to look > > > through > > > > a plastic culture dish. Do you have any experience with live > imaging, > > > > microscopy or dye loading? Have you looked in the literature? There > > are > > > > many papers using Mitotrackers, and in zebrafish. > > > > > > > > Providing a description of your instrumentation, experimental intent > > and > > > > sample preparations when you ask for assistance will avoid a game of > > "20 > > > > Questions". > > > > > > > > Mitotrackers require testing for concentration, incubation times, > > > > temperature, etc. before embarking on any experiment but the > literature > > > > will provide starting points. Overloading can reduce intensity (by > > > > disrupting mitochondrial function), and is a common beginner's > mistake. > > > > try 25 nM to 100 nM, 20-30 min. or shorter. MT Red is brighter, use > > no > > > > more than 25 nM, Deep Red behaves like MT Green, bt possibly a bit > > > voltage > > > > sensitive. > > > > > > > > Digital zoom will not improve signal, it sometimes helps with > > visibility > > > > on the screen. > > > > > > > > Glen MacDonald > > > > Core for Communication Research > > > > Virginia Merrill Bloedel Hearing Research Center > > > > Cellular Morphology Core > > > > Center on Human Development and Disability > > > > Box 357923 > > > > University of Washington > > > > Seattle, WA 98195-7923 USA > > > > (206) 616-4156 > > > > [hidden email] > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > > On Feb 24, 2014, at 3:06 AM, Amandeep Walia <[hidden email]> > wrote: > > > > > > > > > ***** > > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > > ***** > > > > > > > > > > Dear even i am trying to but getting nothing > > > > > > > > > > > > > > > > > > > > On Mon, Feb 24, 2014 at 4:14 PM, Chaitali Banerjee < > > > > > [hidden email]> wrote: > > > > > > > > > >> ***** > > > > >> To join, leave or search the confocal microscopy listserv, go to: > > > > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > >> ***** > > > > >> > > > > >> I am also trying to localize mitochondria in fish macrophages > using > > > > >> mitotracker. What I am getting is greenish tinge of mitotracker > at x > > > 100 > > > > >> objective. Can u please suggest me if there is some software for > > > digital > > > > >> zooming > > > > >> Regards > > > > >> Chaitali > > > > >> > > > > >> > > > > >> On Mon, Feb 24, 2014 at 3:59 PM, Sergey Tauger <[hidden email] > > > > > > wrote: > > > > >> > > > > >>> ***** > > > > >>> To join, leave or search the confocal microscopy listserv, go to: > > > > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > >>> ***** > > > > >>> > > > > >>> Could you please tell what exactly you are going to image and > what > > is > > > > the > > > > >>> label? > > > > >>> Are you going to perform some quantitative imgaing or just make > > sure > > > > that > > > > >>> your > > > > >>> sample really contains mitochondria? What is the cell type? > > > > >>> > > > > >>> Mitchondria are well visible for 1.2 NA objective (TMRE staining, > > > > >>> lymphocytes) and > > > > >>> qualitatively visible while using NA 1 objective if cells are > > > spreaded > > > > >>> (MitoTracker > > > > >>> Green, Vero cells) > > > > >>> > > > > >>> Best > > > > >>> Sergey Tauger > > > > >>> PhD student > > > > >>> Cell motility lab. > > > > >>> Dept. of Biology > > > > >>> Moscow State University > > > > >>> > > > > >> > > > > > > > > > > |
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