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Fluorescent Sensor

Overview of Fluorescent Sensor

Biological sensors are fundamental to current research, from quantifying metabolites under different perturbations to identifying cell states. Many types of sensors exist with a variety of readouts, with one of the most widely used platforms being fluorescent sensors. Fluorescent sensor molecules for imaging cellular molecules have become a hot topic in chemical biology research during the last two decades. A fluorescent sensor is advantageous due to its high sensitivity.

Metabolic targets of fluorescent biosensors in neurons. Fig.1 Metabolic targets of fluorescent biosensors in neurons. (Koveal, 2020)

Application of Fluorescent Sensor in Neuroscience

In neuroscience, the most applied fluorescent sensors are genetically encoded calcium indicators (GECIs), genetically encoded voltage indicators (GEVIs), and pH sensors (pHluorins). Numerous other fluorescent indicators have been applied to study brain cell activity, including sensors for inorganic ions and molecules (chloride, zinc, hydrogen peroxide) and organic signaling molecules and metabolites (glutamate, acetylcholine, ATP, NADH, cyclic nucleotides, glucose, pyruvate, phospholipids). Several sensors that directly monitor enzyme activity have also been developed. These allow the activity of small GTPases, kinases (e.g., protein kinase A), and proteases to be detected. Moreover, GPCR activation can be followed by intra- or intermolecular FRET, either within the heterodimeric G protein or between the GPCR and a binding partner (G protein or arrestin). This approach has been applied to analyze the activation kinetics of metabotropic glutamate receptors, GABAB receptors, and M1 muscarinic acetylcholine receptors.

Sensor Scaffold Fluorescent Protein(s) Excitation Emission Sensor Design Dynamic range: fold change or Δ lifetime Affinity (Kd or KR)
ATP
ATeam1.03 F0F1-ATP synthase, ε subunit (B. subtilis) mseCFP/ mVenus 435 nm (D) 475 nm (D)
527 nm (A)
FRET 2.3-fold (37℃) 3.3 mM
ATeam1.03YEMK F0F1-ATP synthase, ε subunit (B. subtilis) mseCFP/ mVenus 435 nm (D) 475 nm (D)
527 nm (A)
FRET n.r. 1.2 mM (37℃)
2.6 mM (20-22℃)
QUEEN-2m F0F1-ATP synthase, ε subunit (B. subtilis) cp-EGFP 400 nm / 494 nm 513 nm Ratiometric (excitation) >3-fold (25) 2.4 mM
QUEEN-7μ F0F1-ATP synthase, ε subunit (B. PS3) cp-EGFP 400 nm / 494 nm 513 nm Ratiometric (excitation) ~4.3-fold (37℃)
~5-fold (25)
14 μM
7.2 μM
iATPSnFR1.0 F0F1-ATP synthase, ε subunit (B. PS3) cp-SFGFP 488 nm 515 nm Intensity 2-fold (RT) 350 μM
iATPSnFR1.1 F0F1-ATP synthase, ε subunit (B. PS3) cp-SFGFP 488 nm 515 nm Intensity 1.88-fold (RT) 138 μM
ATP:ADP
PercevalHR GlnK, nucleotide binding protein (M. jannaschii) cp-mVenus 482 nm / 455 nm 529 nm Ratiometric (excitation) 4.6-fold (37℃)
~4-fold (RT)
ATP:ADP ≈ 6.1 ATP:ADP ≈ 3.5
NADH
Frex B-Rex, NADH binding protein (B. subtilis) cpYFP 488 nm / 405 nm 525 nm Ratiometric (excitation) ~9.5-fold (RT) 3.7 μM
NADH:NAD+
SoNar T-Rex, NADH binding protein (T. aquaticus) cpYFP 420 nm / 485 nm 528 nm Ratiometric (excitation) ~15-fold (RT) NADH:NAD+ ≈ 1/40
Peredox T-Rex, NADH binding protein (T. aquaticus) cp-T- Sapphire 400 nm 510 nm Intensityb 2.5-fold (35) NADH:NAD+ ≈ 1/90
800 nm (two-photon) 525 nm Lifetime 0.9 ns (35)
0.8 ns (25)
NADH:NAD+ ≈ 1/255 NADH:NAD+ ≈ 1/529
Glucose
FLII12Pglu700μ∆6 MglB, glucose/galactose binding protein (E. coli) eCFP/ Citrine 433 nm (D) 485 nm (D)
528 nm (A)
FRET 1.5-fold (RT) 660 μM
Green Glifon600 MglB, glucose/galactose binding protein (E. coli) Citrine 480 nm 530 nm Intensity ~5-fold (RT) 590 μM
Green Glifon4000 MglB, glucose/galactose binding protein (E. coli) Citrine 480 nm 530 nm Intensity ~6-fold (RT) 3.8 mM
iGlucoSnFR GGBP, glucose/ galactose binding protein (T. thermophilus) cpGFP 485 nm 515 nm Intensity 3.32-fold (RT) 7.7 mM
iGlucoSnFR-TS GGBP, glucose/ galactose binding protein (T. thermophilus) cp-T- Sapphire 790 nm (two-photon) 525 nm Lifetime 0.34 ns (37℃)
0.38 ns (37℃)
2.2 mM
1.8 mMc
Lactate
Laconic LldR, lactate binding transcription regulator (E. coli) mTFP/ Venus 430 nm (D) 480 nm (D)
535 nm (A)
FRET ~1.2-fold (25) Biphasic:
K1 = 8 µM
K2 = 830 µM
Pyruvate
Pyronic PdhR, pyruvate dehydrogenase complex repressor (E. coli) mTFP/ Venus 430 nm (D) 480 nm (D)
535 nm (A)
FRET ~1.24-fold (RT) 107 μM

Table1 Genetically encoded fluorescent biosensors used for the study of neuronal metabolism

Services at Creative Biolabs

With professional expertise, advanced technology platforms, excellent specialists, and rich experience in neuroscience, Creative Biolabs has gained a good reputation in the industry. Years of practice devoted to neuroscience research make us experienced in fluorescent sensors' application in this field. With all these advantages, we are confident in providing customers high-quality relevantly custom productions. If you are interested in applying fluorescent sensors in neuroscience research or have any other inquiries on neuro-based custom productions, please feel free to contact us for more information.

Reference

  1. Koveal, D.; et al. Fluorescent Biosensors for Neuronal Metabolism and the Challenges of Quantitation. Curr Opin Neurobiol. 2020, 63: 111-121.

For Research Use Only. Not For Clinical Use.