Change History for mEos4b

• 2018-11-01 18:12, user: Paezg
  • added protein mEos4b references: Ando et al. (2018)
  • added excerpt Ando et al. (2018): Osmium-resistant probes for fl...
• 2018-11-01 18:13, user: Paezg
  • added protein mEos4b references: Kopek et al. (2017)
• 2018-11-01 18:15, user: Paezg
  • changed protein mEos4b switch_type: o->pc
  • added state transition mEos4b Green -405-> Red
• 2018-11-01 18:22, user: Paezg
  • added excerpt Kopek et al. (2017): We mutated surface residues on...
• 2018-11-01 18:39, user:
  • added protein mEos4b references: Cranfill et al. (2016)
• 2018-11-30 01:32, user:
  • added State mEos4b (Red) brightness: 39.41
  • added State mEos4b (Red) ext_coeff: 55500
  • changed State mEos4b (Green) brightness: 30.34->65.66
  • changed State mEos4b (Green) ext_coeff: 36120->78170
  • added oser measurement OSER: mEos4b in Paez-Segala et al. (2015)
• 2018-11-30 01:37, user: talley
  • changed protein mEos4b seq_validated: False->True
  • changed excerpt Ando et al. (2018): Osmium-resistant probes for fl... content: Osmium-resistant probes for fluorescence and electron microscopy—the fluorescent EosFP derivative mEos4 has recently been described to withstand osmium treatment during EM processing. The mEos4 molecule is therefore still fluorescent in a well-stained resin-embedded sample. In addition, it is compatible with super resolution photoactivated localization microscopy. ->Osmium-resistant probes for fluorescence and electron microscopy—the fluorescent EosFP derivative mEos4 has recently been described to withstand osmium treatment during EM processing. The mEos4 molecule is therefore still fluorescent in a well-stained resin-embedded sample. In addition, it is compatible with super resolution photoactivated localization microscopy.
  • changed excerpt Kopek et al. (2017): We mutated surface residues on... content: We mutated surface residues on mEos2 to remove nucleophilic groups, which are involved in cross-linking with aldehydes and OsO4. This resulted in the selection of two mutants, mEos4a and mEos4b, each with substantially improved resistance to OsO4 fixation and fluorescence properties unchanged from those of the starting scaffold mEos2. These proteins facilitated the development of two protocols: (i) a ‘consecutive-section’ approach in which adjacent ultrathin sections cut from resin are separately split between PALM imaging and EM fixation and imaging; and (ii) a ‘same-section’ approach, in which a single resin-cut section is subjected to both PALM and TEM and/or SEM. In both cases, plastic resin embedding markedly decreases tissue distortion from dehydration and secondary fixation, and additionally improves performance of the specimen under the electron beam. We also found that this protocol is appropriate for use with HPF-FS (e.g., Fig. 1d) or without it (e.g., Fig. 1e). We found ultrastructure preservation to be comparable between the two. ->We mutated surface residues on mEos2 to remove nucleophilic groups, which are involved in cross-linking with aldehydes and OsO4. This resulted in the selection of two mutants, mEos4a and mEos4b, each with substantially improved resistance to OsO4 fixation and fluorescence properties unchanged from those of the starting scaffold mEos2. These proteins facilitated the development of two protocols: (i) a ‘consecutive-section’ approach in which adjacent ultrathin sections cut from resin are separately split between PALM imaging and EM fixation and imaging; and (ii) a ‘same-section’ approach, in which a single resin-cut section is subjected to both PALM and TEM and/or SEM. In both cases, plastic resin embedding markedly decreases tissue distortion from dehydration and secondary fixation, and additionally improves performance of the specimen under the electron beam. We also found that this protocol is appropriate for use with HPF-FS (e.g., Fig. 1d) or without it (e.g., Fig. 1e). We found ultrastructure preservation to be comparable between the two.
• 2018-11-30 01:42, user: talley
  • changed State mEos4b (Red) ex_max: 569->570
  • changed State mEos4b (Green) ex_max: 504->505
• 2019-07-09 00:25, user: talley
  • added protein mEos4b references: De Zitter et al. (2019)
  • removed excerpt Ando et al. (2018): Osmium-resistant probes for fl...
  • removed excerpt Kopek et al. (2017): We mutated surface residues on...
  • added lineage mEos4b
  • added reference De Zitter et al. (2019)
• 2020-09-17 09:34, user: BOURGEOIS
  • added protein mEos4b references: De Zitter et al. (2020)
  • removed reference De Zitter et al. (2019)
  • added reference De Zitter et al. (2020)
• 2021-02-05 14:12, user: jlazar
  • added protein mEos4b references: Myšková et al. (2020)
  • removed reference De Zitter et al. (2020)
• 2024-10-15 13:35, user: kadmon
  • added protein mEos4b references: Maity et al. (2023)
  • changed lineage mEos4b rght: 27->29
  • added reference Maity et al. (2023)

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