LEONARDO DA VINCI PROGRAM

Continuing Education Course
BASICS IN FLUORESCENCE MICROSCOPY
AND FLUOROCHROMES
(November 26-27, 1998)

With the collaboration of

University of Pavia
Department of Animal Biology
Laboratory of Comparative Anatomy
and
Laboratory of Cell Biology
Piazza Botta 10, 27100 Pavia, Italy

Organizers

Prof. Isabel Freitas Laboratorio di Anatomia Comparata Dipartimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506317 fax: + 39 - 0382 - 506406 e-mail: freitas@unipv.it	Prof. Carlo Pellicciari Laboratorio di Istologia ed Antropologia Departimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506420 fax: + 39 - 0382 - 506325 e-mail: pelli@unipv.it
Dr. Heinz Gundlach Carl Zeiss Carl Zeiss Str. 1 D-7347 Oberkochen, Germany phone: + 49 - (0)7364 - 202071 fax: + 49 - (0)7364 - 204013 e-mail: gundlach@zeiss.de
Dr. Thomas Beller Carl Zeiss S.p.A. Viale delle Industrie, 18 20020 Arese (Mi), Italy phone: + 39 - 02 - 937731 fax: + 39 - 02 - 93773301 e-mail: micro@zeiss.it	Dr. Alessandro Borsatti Carl Zeiss S.p.A. Viale delle Industrie, 18 20020 Arese (Mi), Italy phone: + 39 - 02 - 937731 fax. + 39 - 02 - 93773301 e-mail: micro@zeiss.it

Speakers

Dr. Giovanni Bottiroli Centro di Studio per l’Istochimica del C.N.R. Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506412 fax: + 39 - 0382 - 506430 e-mail: botti@dragon.ian.pv.cnr.it	Dr. Heinz Gundlach Carl Zeiss Carl Zeiss Str. 1 D-7347 Oberkochen, Germany phone. + 49 - (0)7364 - 202071 fax: + 49 - (0)7364 - 204013 e-mail: gundlach@zeiss.de
Dr. Giuliano Mazzini Centro di Studio per l’Istochimica del C.N.R. Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506266 fax: + 39 - 0382 - 506430 e-mail:mazzi@dragon.ian.pv.cnr.it	Prof. Sergio Barni Laboratorio di Anatomia Comparata Dipartimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506421 fax: + 39 -0382 - 506406 e-mail: anatcomp@unipv.it
Dr. Luigi Sciola Dipartimento di Scienze Fisiologiche, Biochimiche e Cellulari Università di Sassari Via Muroni 25, 07100 Sassari, Italy phone: + 39 - 079 - 228651 fax: + 39 - 079 - 228615 e-mail: sciola@ssmain.uniss.it

Tutors

Dr. Luigi Sciola Dipartimento di Scienze Fisiologiche, Biochimiche e Cellulari Università di Sassari Via Muroni 25, 07100 Sassari, Italy phone: + 39 - 079 - 228651 fax: + 39 - 079 - 228615 e-mail: sciola@ssmain.uniss.it	Dr. Vittorio Bertone Laboratorio di Anatomia Comparata Dipartimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506317 fax: + 39 - 0382 - 506406 e-mail: bertone@unipv.it
Dr. Patrizia Griffini Laboratorio di Anatomia Comparata Dipartimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506317 fax: + 39 - 0382 - 506406 e-mail: freitas@unipv.it	Dr. Barbara Bono Istituto di Patologia Generale “C. Golgi” Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506334 fax: + 39 - 0382 - 303673 e-mail: golgi@unipv.it
Dr. Simona Fracchiolla Laboratorio di Anatomia Comparata Dipartimento di Biologia Animale Università di Pavia Piazza Botta 10, 27100 Pavia, Italy phone: + 39 - 0382 - 506317 fax: + 39 -0382 - 506406 e-mail: freitas@unipv.it	Dr. Alessandra Spano Dipartimento di Scienze Fisiologiche, Biochimiche e Cellulari Università di Sassari Via Muroni 25, 07100 Sassari, Italy phone: + 39 - 079 - 228651 fax: + 39 - 079 - 228615 e-mail: sciola@ssmain.uniss.it

F I N A L R E P O R T

The continuing education course "Basics in fluorescence microscopy and fluorochromes" held in the 26 and 27th november 1998 in the Department of Animal Biology of the University of Pavia, was attended by 52 researchers in the biomedical field, coming from the Universities of Pavia, Milan and Turin. The theoretical notions imparted in the morning tutorials by Drs. Bottiroli and Mazzini (CNR Center for Histochemistry, Pavia), Prof. Barni (University of Pavia) and Dr. Gundlach (Carl Zeiss, Oberkochen, Germany) were followed in the afternoon by practical workshops during which the partecipants were allowed to observe under the supervision of Dr. Gundlach fluorochromized samples of fixed or supravitally stained cells on microscopes provided by Carl Zeiss SpA, Italy. Multicolor labeling experiments were made on living cells and tissue sections using fluorescence probes provided by Molecular Probes. The partecipants could select single or multibandpass filters in order to obtain the best observation conditions for each sample.
   These two days were preceded by a short training course of three days on supravital fluorescence techniques, held by experts from the Universities of Pavia (Prof. Barni) and Sassari (Drs. Sciola and Spano), and addressed to 5 future tutors of the Leonardo Courses (Drs. Griffini, Bertone, Bono, Fracchiolla and Garda).
   The Course organizers (Prof.s Freitas, Pellicciari and Gundlach) acknowledge that the enormous success of both courses was made possible due to the "back stage" efforts and hard work of the experts Sciola and Spano, the tutors, the technicians from the Laboratory of Cell Biology (Dr. Bottone, Mrs. Veneroni), from Carl Zeiss SpA, Italy (drs. Borsatti and Beller) and the secretaries of the Laboratories of Comparative Anatomy and of Cell Biology (Mr. Sanga and Mrs. Bosini).
   The didactic material was kindly provided by the Gruppo Italiano di Citometria. The digital acquisition and elaboration of the images was made by Drs. Borsatti and Bertone. The Internet advertisements, the electronic elaboration of the booklet with didactic material and the photographic documentation were made by Dr. Bertone.

Selected pictures of the biological samples
>> Click on the images to see at full screen <<

Real color images: scans from color slides

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1. Mast cells at tumor periphery. Acridine Orange (DNA: green; RNA and heparin: orange-red). Single exposure.
2. Fibroblasts supravitally stained with Nile Red (polar lipids: red; neutral lipids: yellow-green) and Hoechst 33342 (DNA: light blue). Double exposure.
3. Fibroblasts supravitally stained with DiOC6 (mitochondria: green) and Hoechst 33342 (DNA: light blue). Double exposure.
4. Fibroblasts supravitally stained with JC1 (low mitochondria membrane potential: green; high potential: red). Single exposure.
5. Fibroblasts supravitally stained with C6-NBD Ceramide (Golgi apparatus: green-yellow). Single exposure.

Pseudo color images: B & W CCD SensiCam PCO camera, Germany; Axiovision software and Axioplan II microscope, Carl Zeiss, Germany

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6. Smooth muscle cells stained with Alexa 488 Phalloidin (actin microfilaments: green) and Hoechst 33342 (DNA). Two single filter sets.
7. Liver section stained with Alexa 488 Phalloidin and Hoechst 33342.
8. Smooth muscle cells supravitally stained with DiOC6 and Hoechst 33342.
9. Suffering smooth muscle cells supravitally stained with Nile Red and Hoechst 33342.

Pictures of the participants....