Seeing through cells : Rapid measurement of intracellular target proteins
Larsson, Per Olof LU and Leiva Eriksson, Nelida LU
(2022)
In Frontiers in Bioengineering and Biotechnology
10.
- Abstract
We have studied a method for making microbial cells transparent by immersing them in a solution with a high refractive index (RI). When the RI of the solution was matching that of the cells, light scattering was greatly diminished (by a factor of up to about 100) and the cell suspension became transparent, facilitating the spectrophotometric determination of intracellular compounds such as hemoglobin. We investigated the properties of several compounds such as sucrose, glycerol, bovine serum albumin, FicollTM, and iodixanol (OptiprepTM), each with advantages and disadvantages. Particularly good overall properties were found for iodixanol at a concentration of around 36% (w/v) and bovine serum albumin at a... (More)
We have studied a method for making microbial cells transparent by immersing them in a solution with a high refractive index (RI). When the RI of the solution was matching that of the cells, light scattering was greatly diminished (by a factor of up to about 100) and the cell suspension became transparent, facilitating the spectrophotometric determination of intracellular compounds such as hemoglobin. We investigated the properties of several compounds such as sucrose, glycerol, bovine serum albumin, FicollTM, and iodixanol (OptiprepTM), each with advantages and disadvantages. Particularly good overall properties were found for iodixanol at a concentration of around 36% (w/v) and bovine serum albumin at a concentration of about 30% (w/v). By using this RI-matching principle the production of intracellular compounds can easily be followed in near real-time during fermentation processes. For example, some conditions for producing plant hemoglobin in Escherichia coli were conveniently determined without the need of any cell disintegration or product purification.
(Less)
- author
- Larsson, Per Olof
LU
and Leiva Eriksson, Nelida
LU
- organization
- publishing date
- 2022
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- BSA, FicollTM, hemoglobin, intracellular proteins, iodixanol, refractive index, transparent cell suspensions
- in
- Frontiers in Bioengineering and Biotechnology
- volume
- 10
- article number
- 996224
- publisher
- Frontiers Media S. A.
- external identifiers
-
- pmid:36263354
- scopus:85140030366
- ISSN
- 2296-4185
- DOI
- 10.3389/fbioe.2022.996224
- language
- English
- LU publication?
- yes
- id
- 0c419740-e95f-4f18-b60b-3de0e3ed2049
- date added to LUP
- 2022-12-13 15:25:35
- date last changed
- 2025-02-21 23:10:03
@article{0c419740-e95f-4f18-b60b-3de0e3ed2049,
abstract = {{<p>We have studied a method for making microbial cells transparent by immersing them in a solution with a high refractive index (RI). When the RI of the solution was matching that of the cells, light scattering was greatly diminished (by a factor of up to about 100) and the cell suspension became transparent, facilitating the spectrophotometric determination of intracellular compounds such as hemoglobin. We investigated the properties of several compounds such as sucrose, glycerol, bovine serum albumin, Ficoll<sup>TM,</sup> and iodixanol (Optiprep<sup>TM</sup>), each with advantages and disadvantages. Particularly good overall properties were found for iodixanol at a concentration of around 36% (w/v) and bovine serum albumin at a concentration of about 30% (w/v). By using this RI-matching principle the production of intracellular compounds can easily be followed in near real-time during fermentation processes. For example, some conditions for producing plant hemoglobin in Escherichia coli were conveniently determined without the need of any cell disintegration or product purification.</p>}},
author = {{Larsson, Per Olof and Leiva Eriksson, Nelida}},
issn = {{2296-4185}},
keywords = {{BSA; FicollTM; hemoglobin; intracellular proteins; iodixanol; refractive index; transparent cell suspensions}},
language = {{eng}},
publisher = {{Frontiers Media S. A.}},
series = {{Frontiers in Bioengineering and Biotechnology}},
title = {{Seeing through cells : Rapid measurement of intracellular target proteins}},
url = {{http://dx.doi.org/10.3389/fbioe.2022.996224}},
doi = {{10.3389/fbioe.2022.996224}},
volume = {{10}},
year = {{2022}},
}