magnification for an in deep observation discrete of cell sections and organelles ultrastructure.
4. Take pictures of the sections of interest using a SIS Megaview
III digital camera.
4 Notes
1. Paraformaldehyde and formaldehyde are toxic by inhalation,
ingestion and skin absorption and corrosive on skin contact.
Wear gloves and use a cupboard.
2. It should be protected from CO 2 since produces a precipitate.
It should not be used if it becomes cloudy.
3. Lead nitrate is toxic by inhalation and harmful by ingestion and
skin absorption. Avoid breathing and use gloves.
4. Uranyl acetate is very toxic by ingestion and inhalation. Wear
gloves and prepare the solution in a fume cup-board.
5. Centrifuge both stains before using (put an aliquot of 0.5 mL
in an Eppendorf tube) and centrifuge at maximum speed.
6. Uranyl acetate should be made always before lead staining.
7. Uranyl acetate produce an undesired precipitate on exposure to
light, therefore grids should be stained and stored in darkness
afterward.
8. Both stains are harmful by inhalation and ingestion. Avoid
breathing dust and wear gloves. In case of spillage wash with
abundant water.
9. Leave samples to dry 24 h before TEM visualization. Store in
the dark.
Acknowledgments
Financial support by research grants AGL-2015-70218 (Ministerio
Economı ´a y Competitividad (Spain), and FEDEGENT/2018/007
(Generalitat Valenciana, Spain) are gratefully acknowledged. MJR
and LZ are members of Eurocaroten (COST_Action CA15136)
and CaRed (Spanish Carotenoid Network, BIO2015-71703REDT and BIO2017-90877-REDT).
References
1. Rodriguez-Concepcion M, Avalos J, Bonet
ML, Boronat A, Gomez-Gomez L, HorneroMendez D, Limon MC, Mele ´ndez-Martı ´nez
AJ, Olmedilla-Alonso B, Palou A, Ribot J,
Rodrigo MJ, Zacarias L, Changfu Z (2018) A
global perspective on carotenoids: metabolism,
biotechnology, and benefits for nutrition and
health. Prog Lipid Res 70:62–93
242
Joanna Lado et al.
4. Take pictures of the sections of interest using a SIS Megaview
III digital camera.
4 Notes
1. Paraformaldehyde and formaldehyde are toxic by inhalation,
ingestion and skin absorption and corrosive on skin contact.
Wear gloves and use a cupboard.
2. It should be protected from CO 2 since produces a precipitate.
It should not be used if it becomes cloudy.
3. Lead nitrate is toxic by inhalation and harmful by ingestion and
skin absorption. Avoid breathing and use gloves.
4. Uranyl acetate is very toxic by ingestion and inhalation. Wear
gloves and prepare the solution in a fume cup-board.
5. Centrifuge both stains before using (put an aliquot of 0.5 mL
in an Eppendorf tube) and centrifuge at maximum speed.
6. Uranyl acetate should be made always before lead staining.
7. Uranyl acetate produce an undesired precipitate on exposure to
light, therefore grids should be stained and stored in darkness
afterward.
8. Both stains are harmful by inhalation and ingestion. Avoid
breathing dust and wear gloves. In case of spillage wash with
abundant water.
9. Leave samples to dry 24 h before TEM visualization. Store in
the dark.
Acknowledgments
Financial support by research grants AGL-2015-70218 (Ministerio
Economı ´a y Competitividad (Spain), and FEDEGENT/2018/007
(Generalitat Valenciana, Spain) are gratefully acknowledged. MJR
and LZ are members of Eurocaroten (COST_Action CA15136)
and CaRed (Spanish Carotenoid Network, BIO2015-71703REDT and BIO2017-90877-REDT).
References
1. Rodriguez-Concepcion M, Avalos J, Bonet
ML, Boronat A, Gomez-Gomez L, HorneroMendez D, Limon MC, Mele ´ndez-Martı ´nez
AJ, Olmedilla-Alonso B, Palou A, Ribot J,
Rodrigo MJ, Zacarias L, Changfu Z (2018) A
global perspective on carotenoids: metabolism,
biotechnology, and benefits for nutrition and
health. Prog Lipid Res 70:62–93
242
Joanna Lado et al.