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Table 8 Fluorescence spectroscopy methods

From: Lipids detection and quantification in oleaginous microorganisms: an overview of the current state of the art

Method Microorganism Range Sample preparation Correlation with method Internal Standard Excitation/emission (nm) Dye concentration Reference
Nile Red Amphora coffeaformis, Navicula sp., Tropidoneis sp. n/a Whole cells Gravimetric, GC Exc. 488–525
Em. 570–600
1 μg /mL/acetone [37]
Nile Red (high-throughput microplate assay with mild bleach solution addition) 3 Chlorella species 0.75–40 μg Lipid extracts suspended in chloroform; whole cells; extracted lipids (25 μL diluted in isopropanol); cells (200 μL OD600 0.35–1.0) GC/MS, gravimetric, TAG determination with TLC Corn oil Exc. 530
Em. 575
4 μg/mL/ DMSO [165]
Nile Red Lipomyces starkeyi, Rhodosporidium toluroides, Cryptococcus curvatus,3 Mortierella species 2–5000 μg lipid/mL broth Whole cells, 100 μL culture broth suspended in 2 mL buffer GC Exc. 400
Em. 565–585
0.24–0.47 μg/mL/acetone [179]
Nile Red (High-throughput microplate assay) 9 microalgal species 2.0–20.0 μg/mL Whole cells, 3 × 106 cells/mL, OD750 = 0.06 Gravimetric Triolein Exc. 530
Em. 575
0.5 μg/mL /DMSO [180]
Nile Red Nannochloropsis sp. n/a Whole cells, 3 mL of culture Exc. 486
Em. 570
0.26 μM/acetone [156]
Nile Red Skeletonema marinoi, Chaetoceros socialis, Alexandrium minutum 0.12–12 μg/mL Whole cells, 2 mL resuspended in isopropanols and internal standard Triolein Exc .547
Em.580
0.25 μg/mL [161]
Nile Red (High-throughput microplate) Saccharomyces cerevisiae, Rhodosporidium diobovatum, Pichia manshurica, Rodosporidium paledigenum, Cryptococcus victoriae n/a Whole cells, 250 μL at OD600 = 1 in 300 μL total volume Gravimetric Exc.530
Em. 590
5 μg/mL /DMSO [162]
Nile Red (High-throuput microplate assay, two-step, microwave assisted) Pseudochlorococcum sp., Scenedesmus dimorphus, Chlorella zofigiensis n/a Whole cells, 5–10 μL algal cells Exc. 490
Em. 580
1 μg/mL /DMSO and acetone [159]
Nile Red (High-throughput assay) Auxenochlorella protothecoides, Chlorella vulgaris, Scenedesmus dimorphus, Scenedesmus obliquus n/a Whole cells, 5 g/L dry weight, 10 μL algal suspension in 120 μL total volume Exc. 530
Em. 604
10 μg/mL /ethanol [171]
Nile Red A. protothecoides Up to 70% dry weight Whole cells, 1.6 mL SPV Exc. 495
Em. 540–750
6 μg /mL/ DMSO [181]
Nile Red (cell treatment with EDTA) Tetraselmis suecica, 6 microalgal stains Whole cells, 250 μL Exc. 530
Em. 590
1.3 μg/mL / acetone [164]
Nile Red Nannochloropsis sp. Whole cells, 2 mL    Exc. 480
Em. 575
0.3 and 0.7 μg/mL
for glycerol and DMSO, respectively
[160]
Nile Red (combining ultrasonic treatment and 3D excitation emission matrix fluorescence spectroscopy) Scenedesmus sp. n/a Whole cells, 5 mL Gravimetric Exc. 530
Em. 568
1.5 mg/mL /acetone [163]
Nile Red Nannochloropsis oculata, Tisochrysis lutea n/a Whole cells, OD750 = 0.2 Gravimetric Exc. 570
Em. 670
1.67 or 3.33 μg/mL/ glycerol [158]
BODIPY 505/515 C. vulgaris, Dunaliella primolecta, Chaetoceros calcitrans n/a Whole cells, 106 cells/mL Nile Red Exc. 480
Em. 590
0.067 μg/mL/DMSO [168]
AC-202 Chlamydomonas reinhardtii, Phaeodactylum tricornutum n/a Whole cells, 1 mL Nile Red, BODIPY Exc. 395
Em. BP445/50
10 μM/DMSO [172]
  1. n/a Not available