Project developed by

Dr. Jane Elizabeth Kraus (kraus@usp.br)

&

Juliana Pisaneschi (jupisa@ib.usp.br)

 

Material and Methods

Work Team and acknowledgements

References

Financial Support


Material and Methods

The plants used in this small atlas were selected because they were readily available. Chosen due to its easy preparation, temporary histological slides were mounted with hand made sections. These histological slides are easily prepared by students in different classes of Botany.

Monocotyledons

Root: orchid (Epidendrum fulgens, Orchidaceae)
Stem: daylily (Hemerocallis flava, Liliaceae)
Leaf: lemon-grass (Cymbopogon citratus, Poaceae)

Dicotyledons

Root: wild strawberry plant (Rubus rosaefolius, Rosaceae)
Stem: castor oil plant (Ricinus communis, Euphorbiaceae)
Leaf: coffee plant (Coffea arabica, Rubiaceae)

a) Sample conservation

Roots, stems and leaves were individually separated, divided in smaller portions and maintained in flasks with 70% ethanol for conservation.

b) Sample fragmentation

Cylindrical organs, like roots and stems, were sectioned in 2-4cm fragments in lenght. Leaf blades were cut into 0.25cm2 pieces (taken from their middle portion, enclosing the major vein).

c) Sample sectioning

In the case of soft tissues or small plant parts, the use of a support was necessary to make the sectioning easier. The plant fragment used in the study was placed between two pieces of cork or styrofoam. From this
fragment transverse hand made sections were obtained with the help of razor blades. These sections were placed in a watch glass containing water.

d) Clarification of the histological sections

Using a thin brush, histological sections were transferred to another watch glass containing 50% commercial household bleach until they became whitish (3-5 minutes, depending on the material). The bleach was removed with a Pasteur pipette and the sections were washed carefully 2-3 times with water, until the preparation became odorless.

e) Staining of the histological sections

The sections were transferred with a brush to another watch glass containing a mixture of 1% astra blue and 1% safranine (9:1, v/v). The sections were stained for less than a minute (variable time, depending on the material). These stained histological sections were transferred with a brush, to another watch glass containing water. The sections were washed carefully 2-3 times for the removal of excess stain.

f) Slide mounting

Histological sections were placed on slides containing 50% glycerin drops. A cover glass was laid over each section and sealed with transparent nail polish on the borders.

g) Photomicrographs

Photomicrographs were taken from the histological sections using color films (ASA 100) and a blue filter.

h) Diagrams

Diagrams were made from the histological sections using a projection microscope.

i) Scanning of the photomicrographs and diagrams

The diagrams and photomicrographs were scanned with a table top scanner.

h) HTML Page

Page development was carried out with HTML. Each photo was slightly modified with Paint Shop Pro (Jascon Inc.). These two images were overlaid on an applet using JavaScript Language. A simple text editor (Notepad) was used for HTML structure.


 

WORK TEAM

Hypertext programming:

Juliana Pisaneschi

Diagrams:

Makoto Tanoue

Photomicrographs:

Jane Elizabeth Kraus

Histological preparations:

Adriano M. de Castro
Cláudia Vecchi
Delmira da Costa e Silva
Patricia Borges Pita

Concepts:

Jane Elizabeth Kraus

English:

Dr Peter Gibbs
Plant Sciences Laboratory, School of Biology, University of St. Andrews, Scotland, United Kingdom

Dr Wagner de Melo Ferreira
Federal University of Tocantins, Porto Nacional (TO) Brazil

 

 

ACKNOWLEDGMENTS

Claudia Bozzo
Daniel Germano
Daniel Lobato Duclós
Eduardo Ascenço Reis
Ernst Weber
George Uemura
Instituto de Biociências


REFERENCES

 

CUTTER, E. G. 1978. Plant Anatomy: cells and tissues, 2nd ed., Edward Arnold Ltd., London, 315 p.  

CUTTER, E. G. 1980. Plant Anatomy: experiment and interpretation, Edward Arnold Ltd., East Kilbride, 343 p.  

ESAU, K. 1974. Anatomia das plantas com sementes. Ed. Blücher Ltda, São Paulo, 293p.

GEMTCHÚJNICOV, I. D. 1976. Manual de taxonomia vegetal: plantas agrícolas, ornamentais e medicinais. Ed. Agronômica Ceres, São Paulo, 368 p.

FAHN, A. 1990. Plant Anatomy, 4th ed., Pergamon Press, Oxford, 587 p.  

KRAUS, J. E., RECHULKI, R. K. & MARCHINI, S. 1994. Guia para o jardim do Departamento de Botânica do Instituto de Biociências, Instituto de Biociências da USP, São Paulo, 57 p.  

KRAUS, J. E. & ARDUIN, M. 1997. Manual básico de métodos em morfologia vegetal, EDUR, Seropédica, 198 p.  

MAUSETH, J. D. 1991. Botany: an introduction to plant biology, Saunders College, Philadelphia, 800 p.  

MAUSETH, J. D. 1988. Plant anatomy, The Benjamin/Cummings Publish. Co, Menlo Park, 560 p.  

RAMALHO, J. A. 1997. HTML Avançado versão 3.2, Makron Books, São Paulo, 659 p.  

RAVEN, P. H., EVERT, R. F. & EICHORN, S. E. 1996. Biologia vegetal, 5th ed., Ed. Guanabara Koogan, Rio de Janeiro, 728 p.  

SCAVONE, O. & PANIZZA, S. 1980. Plantas tóxicas, CODAC-USP, São Paulo, 110 p.  

THOMAS, M. D., 1997. Programando em Java para a Internet, Makron Books, São Paulo, 665 p.


FINANCIAL SUPPORT

Pró-Reitoria de Graduação (USP)

Pró-Reitoria de Pós-Graduação (USP)


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