Mitochondria: Fluorescence Microscopy vs. SEM
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Engstrom, Lars |
Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello - Since the first time I fluorescently stained mitochondria, I have always wondered why the structure of mitochondria look so different from the classical SEM images. Can someone please explain? Thank you for your time. -Lars |
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Lemasters, John J. |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Different how? I don't see a difference. -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 2:16 PM To: [hidden email] Subject: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello - Since the first time I fluorescently stained mitochondria, I have always wondered why the structure of mitochondria look so different from the classical SEM images. Can someone please explain? Thank you for your time. -Lars |
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Engstrom, Lars |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Maybe it is merely a magnification/resolution or optical slice difference. The cartoon kidney bean-like textbook figures closely approximate the SEM images where as fluorescent images surprised me at their length and "wormy-ness" (I know very scientific). I gave a presentation about DNA, Cells and Art to my daughter's 5th grade class. I was showing some images I have taken of mitochondria and couldn't definitively explain why they looked so different from the textbook images. -Lars -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Lemasters, John J. Sent: Monday, September 26, 2011 11:25 AM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Different how? I don't see a difference. -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 2:16 PM To: [hidden email] Subject: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello - Since the first time I fluorescently stained mitochondria, I have always wondered why the structure of mitochondria look so different from the classical SEM images. Can someone please explain? Thank you for your time. -Lars |
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James Denegre |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Lars, The textbook images are an inaccurate representation of the 3D nature of mitochondria. Mitochondria usually exist in reticulated networks, not as individual "beans" as they are often illustrated in text books. The "bean" image is a misrepresentation of SEM data; the SEM data are from cross-sections of arms of the reticulated network, and do not represent the entire structure. If an extensive serial reconstruction were performed on SEM data a reticulated network would be appear. Reticulated networks of mitochondria are well-illustrated in: Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. 4Pi microscopy reveals an impaired three-dimensional mitochondrial network of pancreatic islet beta-cells, an experimental model of type-2 diabetes. Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, Lessard M, Bewersdorf J, Jezek P, Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. Mitochondria, redox signaling and axis specification in metazoan embryos Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. Regards, Jim James Denegre, Ph.D. Senior Manager Imaging Sciences The Jackson Laboratory Bar Harbor, ME 04609 207.288.6648 On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Maybe it is merely a magnification/resolution or optical slice > difference. The cartoon kidney bean-like textbook figures closely > approximate the SEM images where as fluorescent images surprised me at > their length and "wormy-ness" (I know very scientific). > > I gave a presentation about DNA, Cells and Art to my daughter's 5th > grade class. I was showing some images I have taken of mitochondria and > couldn't definitively explain why they looked so different from the > textbook images. > > -Lars > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Lemasters, John J. > Sent: Monday, September 26, 2011 11:25 AM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair > Director, Center for Cell Death, Injury & Regeneration > Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & > Molecular Biology > Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have always > wondered why the structure of mitochondria look so different from the > classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars |
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Engstrom, Lars |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That was the answer I was looking for! Thanks. -Lars -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of James Denegre Sent: Monday, September 26, 2011 12:42 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Lars, The textbook images are an inaccurate representation of the 3D nature of mitochondria. Mitochondria usually exist in reticulated networks, not as individual "beans" as they are often illustrated in text books. The "bean" image is a misrepresentation of SEM data; the SEM data are from cross-sections of arms of the reticulated network, and do not represent the entire structure. If an extensive serial reconstruction were performed on SEM data a reticulated network would be appear. Reticulated networks of mitochondria are well-illustrated in: Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. 4Pi microscopy reveals an impaired three-dimensional mitochondrial network of pancreatic islet beta-cells, an experimental model of type-2 diabetes. Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, Lessard M, Bewersdorf J, Jezek P, Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. Mitochondria, redox signaling and axis specification in metazoan embryos Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. Regards, Jim James Denegre, Ph.D. Senior Manager Imaging Sciences The Jackson Laboratory Bar Harbor, ME 04609 207.288.6648 On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Maybe it is merely a magnification/resolution or optical slice > difference. The cartoon kidney bean-like textbook figures closely > approximate the SEM images where as fluorescent images surprised me at > their length and "wormy-ness" (I know very scientific). > > I gave a presentation about DNA, Cells and Art to my daughter's 5th > grade class. I was showing some images I have taken of mitochondria and > couldn't definitively explain why they looked so different from the > textbook images. > > -Lars > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Lemasters, John J. > Sent: Monday, September 26, 2011 11:25 AM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair > Director, Center for Cell Death, Injury & Regeneration > Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & > Molecular Biology > Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have always > wondered why the structure of mitochondria look so different from the > classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars |
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Lemasters, John J. |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** This is purely a sectioning issue. Make thin sections of a bowl of spaghetti and you think you have a bowl of rice. John -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 3:45 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That was the answer I was looking for! Thanks. -Lars -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of James Denegre Sent: Monday, September 26, 2011 12:42 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Lars, The textbook images are an inaccurate representation of the 3D nature of mitochondria. Mitochondria usually exist in reticulated networks, not as individual "beans" as they are often illustrated in text books. The "bean" image is a misrepresentation of SEM data; the SEM data are from cross-sections of arms of the reticulated network, and do not represent the entire structure. If an extensive serial reconstruction were performed on SEM data a reticulated network would be appear. Reticulated networks of mitochondria are well-illustrated in: Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. 4Pi microscopy reveals an impaired three-dimensional mitochondrial network of pancreatic islet beta-cells, an experimental model of type-2 diabetes. Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, Lessard M, Bewersdorf J, Jezek P, Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. Mitochondria, redox signaling and axis specification in metazoan embryos Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. Regards, Jim James Denegre, Ph.D. Senior Manager Imaging Sciences The Jackson Laboratory Bar Harbor, ME 04609 207.288.6648 On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Maybe it is merely a magnification/resolution or optical slice > difference. The cartoon kidney bean-like textbook figures closely > approximate the SEM images where as fluorescent images surprised me at > their length and "wormy-ness" (I know very scientific). > > I gave a presentation about DNA, Cells and Art to my daughter's 5th > grade class. I was showing some images I have taken of mitochondria > and couldn't definitively explain why they looked so different from > the textbook images. > > -Lars > > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Lemasters, John J. > Sent: Monday, September 26, 2011 11:25 AM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair Director, > Center for Cell Death, Injury & Regeneration Departments of > Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular > Biology Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have > always wondered why the structure of mitochondria look so different > from the classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars |
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Engstrom, Lars |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** John, are you now saying you do see a difference? :) -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Lemasters, John J. Sent: Monday, September 26, 2011 12:52 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** This is purely a sectioning issue. Make thin sections of a bowl of spaghetti and you think you have a bowl of rice. John -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 3:45 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That was the answer I was looking for! Thanks. -Lars -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of James Denegre Sent: Monday, September 26, 2011 12:42 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Lars, The textbook images are an inaccurate representation of the 3D nature of mitochondria. Mitochondria usually exist in reticulated networks, not as individual "beans" as they are often illustrated in text books. The "bean" image is a misrepresentation of SEM data; the SEM data are from cross-sections of arms of the reticulated network, and do not represent the entire structure. If an extensive serial reconstruction were performed on SEM data a reticulated network would be appear. Reticulated networks of mitochondria are well-illustrated in: Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. 4Pi microscopy reveals an impaired three-dimensional mitochondrial network of pancreatic islet beta-cells, an experimental model of type-2 diabetes. Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, Lessard M, Bewersdorf J, Jezek P, Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. Mitochondria, redox signaling and axis specification in metazoan embryos Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. Regards, Jim James Denegre, Ph.D. Senior Manager Imaging Sciences The Jackson Laboratory Bar Harbor, ME 04609 207.288.6648 On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Maybe it is merely a magnification/resolution or optical slice > difference. The cartoon kidney bean-like textbook figures closely > approximate the SEM images where as fluorescent images surprised me at > their length and "wormy-ness" (I know very scientific). > > I gave a presentation about DNA, Cells and Art to my daughter's 5th > grade class. I was showing some images I have taken of mitochondria > and couldn't definitively explain why they looked so different from > the textbook images. > > -Lars > > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Lemasters, John J. > Sent: Monday, September 26, 2011 11:25 AM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair Director, > Center for Cell Death, Injury & Regeneration Departments of > Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular > Biology Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have > always wondered why the structure of mitochondria look so different > from the classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars |
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Lemasters, John J. |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** It's the same thing that to the inexperienced eye seems different. Also, be sure you are not comparing apples and oranges. Mitochondria of most cells in culture are filamentous but mitochondria in liver, heart and brain generally are not. Textbooks show TEMs (not SEMs) of tissues generally, not cultured cells. -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 4:02 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** John, are you now saying you do see a difference? :) -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Lemasters, John J. Sent: Monday, September 26, 2011 12:52 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** This is purely a sectioning issue. Make thin sections of a bowl of spaghetti and you think you have a bowl of rice. John -- John J. Lemasters, MD, PhD Professor and GlaxoSmithKline Distinguished Endowed Chair Director, Center for Cell Death, Injury & Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina DD504 Drug Discovery Building 70 President Street, MSC 140 Charleston, SC 29425 Office: 843-876-2360 Lab: 843-876-2354 Fax: 843-876-2353 Email: [hidden email] http://academicdepartments.musc.edu/ccdir -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Engstrom, Lars Sent: Monday, September 26, 2011 3:45 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That was the answer I was looking for! Thanks. -Lars -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of James Denegre Sent: Monday, September 26, 2011 12:42 PM To: [hidden email] Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Lars, The textbook images are an inaccurate representation of the 3D nature of mitochondria. Mitochondria usually exist in reticulated networks, not as individual "beans" as they are often illustrated in text books. The "bean" image is a misrepresentation of SEM data; the SEM data are from cross-sections of arms of the reticulated network, and do not represent the entire structure. If an extensive serial reconstruction were performed on SEM data a reticulated network would be appear. Reticulated networks of mitochondria are well-illustrated in: Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. 4Pi microscopy reveals an impaired three-dimensional mitochondrial network of pancreatic islet beta-cells, an experimental model of type-2 diabetes. Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, Lessard M, Bewersdorf J, Jezek P, Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. Mitochondria, redox signaling and axis specification in metazoan embryos Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. Regards, Jim James Denegre, Ph.D. Senior Manager Imaging Sciences The Jackson Laboratory Bar Harbor, ME 04609 207.288.6648 On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Maybe it is merely a magnification/resolution or optical slice > difference. The cartoon kidney bean-like textbook figures closely > approximate the SEM images where as fluorescent images surprised me at > their length and "wormy-ness" (I know very scientific). > > I gave a presentation about DNA, Cells and Art to my daughter's 5th > grade class. I was showing some images I have taken of mitochondria > and couldn't definitively explain why they looked so different from > the textbook images. > > -Lars > > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Lemasters, John J. > Sent: Monday, September 26, 2011 11:25 AM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair Director, > Center for Cell Death, Injury & Regeneration Departments of > Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular > Biology Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have > always wondered why the structure of mitochondria look so different > from the classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars |
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John Oreopoulos |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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In reply to this post by Engstrom, Lars
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Although not SEM, this paper (see Figure 3) has a very direct comparison between fluorescence (super-resolution) and TEM images of mitochondria: BETZIG, E., PATTERSON, G.H., SOUGRAT, R., LINDWASSER, O.W., OLENYCH, S., BONIFACINO, J.S., DAVIDSON, M.W., LIPPINCOTT-SCHWARTZ, J. & HESS, H.F. (2006). Imaging intracellular fluorescent proteins at nanometer resolution. Science 313(5793), 1642-1645. John Oreopoulos Research Assistant Spectral Applied Research Richmond Hill, Ontario Canada www.spectral.ca On 2011-09-26, at 3:45 PM, Engstrom, Lars wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > That was the answer I was looking for! > Thanks. > -Lars > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of James Denegre > Sent: Monday, September 26, 2011 12:42 PM > To: [hidden email] > Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Lars, > > The textbook images are an inaccurate representation of the 3D nature of > mitochondria. Mitochondria usually exist in reticulated networks, not as > individual "beans" as they are often illustrated in text books. > > The "bean" image is a misrepresentation of SEM data; the SEM data are from > cross-sections of arms of the reticulated network, and do not represent the > entire structure. If an extensive serial reconstruction were performed on > SEM data a reticulated network would be appear. > > Reticulated networks of mitochondria are well-illustrated in: > > Mitochondrial oxidative phosphorylation and energetic status are reflected > by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by > 4Pi microscopy.Plecitá-Hlavatá L, Lessard M, Santorová J, Bewersdorf J, > Jezek P. Biochim Biophys Acta. 2008 Jul-Aug;1777(7-8):834-46. > > > 4Pi microscopy reveals an impaired three-dimensional mitochondrial network > of pancreatic islet beta-cells, an experimental model of type-2 diabetes. > Dlasková A, Spacek T, Santorová J, Plecitá-Hlavatá L, Berková Z, Saudek F, > Lessard M, Bewersdorf J, Jezek P, > Biochim Biophys Acta. 2010 Jun-Jul;1797(6-7):1327-41. > > Mitochondria, redox signaling and axis specification in metazoan embryos > Coffman J, Denegre J, Developmental Biology 308 (2007) 266 280. > > > Regards, > > Jim > > James Denegre, Ph.D. > Senior Manager > Imaging Sciences > The Jackson Laboratory > Bar Harbor, ME 04609 > 207.288.6648 > > > > > On 9/26/11 2:48 PM, "Engstrom, Lars" <[hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Maybe it is merely a magnification/resolution or optical slice >> difference. The cartoon kidney bean-like textbook figures closely >> approximate the SEM images where as fluorescent images surprised me at >> their length and "wormy-ness" (I know very scientific). >> >> I gave a presentation about DNA, Cells and Art to my daughter's 5th >> grade class. I was showing some images I have taken of mitochondria and >> couldn't definitively explain why they looked so different from the >> textbook images. >> >> -Lars >> >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:[hidden email]] >> On Behalf Of Lemasters, John J. >> Sent: Monday, September 26, 2011 11:25 AM >> To: [hidden email] >> Subject: Re: Mitochondria: Fluorescence Microscopy vs. SEM >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Different how? I don't see a difference. >> >> -- >> John J. Lemasters, MD, PhD >> Professor and GlaxoSmithKline Distinguished Endowed Chair >> Director, Center for Cell Death, Injury & Regeneration >> Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & >> Molecular Biology >> Medical University of South Carolina >> DD504 Drug Discovery Building >> 70 President Street, MSC 140 >> Charleston, SC 29425 >> >> Office: 843-876-2360 >> Lab: 843-876-2354 >> Fax: 843-876-2353 >> Email: [hidden email] >> http://academicdepartments.musc.edu/ccdir >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:[hidden email]] >> On Behalf Of Engstrom, Lars >> Sent: Monday, September 26, 2011 2:16 PM >> To: [hidden email] >> Subject: Mitochondria: Fluorescence Microscopy vs. SEM >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Hello - >> >> Since the first time I fluorescently stained mitochondria, I have always >> wondered why the structure of mitochondria look so different from the >> classical SEM images. Can someone please explain? >> >> >> >> Thank you for your time. >> >> -Lars |
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Iain Johnson |
Re: Mitochondria: Fluorescence Microscopy vs. SEM
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In reply to this post by Lemasters, John J.
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** How the fluorescent staining is done can also be a factor. Most mitochondrial staining dyes such as TMRE and MitoTracker are dependent (often in a nonlinear way) on respiratory potential. So they produce a functional image that is conditionally related to the underlying structure. A less biased structural image is produced by fluorescent protein fusions or labeled antibodies targeted to uniformly expressed mitochondrial proteins. Iain On Mon, Sep 26, 2011 at 11:24 AM, Lemasters, John J. <[hidden email]>wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Different how? I don't see a difference. > > -- > John J. Lemasters, MD, PhD > Professor and GlaxoSmithKline Distinguished Endowed Chair > Director, Center for Cell Death, Injury & Regeneration > Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & > Molecular Biology > Medical University of South Carolina > DD504 Drug Discovery Building > 70 President Street, MSC 140 > Charleston, SC 29425 > > Office: 843-876-2360 > Lab: 843-876-2354 > Fax: 843-876-2353 > Email: [hidden email] > http://academicdepartments.musc.edu/ccdir > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Engstrom, Lars > Sent: Monday, September 26, 2011 2:16 PM > To: [hidden email] > Subject: Mitochondria: Fluorescence Microscopy vs. SEM > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello - > > Since the first time I fluorescently stained mitochondria, I have always > wondered why the structure of mitochondria look so different from the > classical SEM images. Can someone please explain? > > > > Thank you for your time. > > -Lars > |
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