Literature summary of live-cell applications
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Amytracker molecules are small, non-toxic, and live-cell compatible. They become fluorescent upon binding to protein aggregates, enabling direct visualization and tracking of protein aggregation over time. Their use in live-cell research has been demonstrated in numerous publications, several of which are summarized below.
| Publication | Cell type | Amytracker variant | Target | Staining conditions | Detection method |
|---|---|---|---|---|---|
| Ariyath 2026 | MC65 cells (neuroblastoma) | Amytracker 680 | Aβ aggregates | 30 min incubation at 37°C, in Opti-MEM. Excess dye was removed with three washes. Fluorescence readings were collected daily for eight days. Fluorescence microscopy on day 6. | Microplate reader & fluorescence microscopy |
| Eltom 2024 | hiPSC-derived astrocytes | Amytracker 680 | Tau fibrils | Cells were exposed to brain-derived AD fibrils at a concentration of 25 mg starting tissue/ml in culture medium for 3 days. The cells were then washed 3 × 5 min with PBS and subsequently stained with Amytracker 680. Following a 30-minute incubation, the cells were washed 3 × 5 min with PBS and tau free culture medium was added. | Time-lapse microscopy, confocal fluorescence microscopy |
| Raymundo 2024 | HaCaT cells | Amytracker 630 | Amyloid-like aggregates | Not specified in the publication. | Confocal fluorescence microscopy |
| Li 2024 | Macrophages | Amytracker 680 | Aβ fibrils | The sonicated Aβ fibrils (4 μM total monomer concentration) were first tagged with a 1:1000 dilution of Amytracker 680; this was then delivered to the macrophage surface. | Fluorescence microscopy |
| Pinzi 2024 | Primary neurons from mouse dorsal root ganglia (DRG) | Amytracker 680 | Amyloid fibrils | Serum-reduced DMEM containing NGF and without phenol red one day prior to live imaging. Amytracker 680 was diluted 1:500 in the culture medium. After 30 minutes of incubation, cells were imaged with a 453-nm laser. | Fluorescence microscopy |
| Chandhok 2023 | MCF-7 cells (Breast cancer cell line) | Amytracker 680 | Aβ (1–42) aggregates | Amytracker dye (red) was added to the media of live cells and images were captured. MCF-7 cells (ATCC) were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) (high glucose) which was supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin–streptomycin, which were incubated at 37 °C and 5% carbon dioxide. | Confocal fluorescence microscopy |
| Juliani 2023 | HEK293 cells | Amytracker 680 | Prion aggregates | AmyTracker 680 (Ebba Biotech) labeling of live cells was performed with 1:1000 of the dye in Opti-MEM for 30 min (37°C, 5% CO2) followed by replacing with fresh medium. | Fluorescence microscopy |
| Hochmair 2022 | HEK sensor cells | Amytracker 680 | Tau aggregates | Likely live-cell staining, but fixation status not explicitly stated. | Fluorescence microscopy |
| Choi 2022 | SNCA-A53T hiPSC-derived neurons | Amytracker 540 | α-Syn aggregates | Cells were washed twice and loaded with 1% Amytracker for 20 min. Room temperature HBSS (Hank’s Balanced Salt Solution) was used as a recording buffer. | Confocal fluorescence microscopy |
| Morten 2022 | HEK293 cells | Amytracker 630 | α-Syn, tau, Aβ aggregates | Live-cell imaging of these cells incubated with 1 μM aggregates was conducted in FluoroBrite Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum. | TIRF microscope |
| Hofbauer 2021 | Macrophages | Amytracker 480 | Amyloid fibrils | Cells were washed with PBS and incubated with the AmyTracker 480 reagent (1:1000, 1 h) at 37°C and 5% CO2. As assessed by flow cytometry, an increase in amyloids was defined as increase in fluorescence. | Flow cytometry |