Lysosome Function Analysis - Selection Guide for Detection and Imaging Reagent / Probe / Kit

Lysosomes are essential for maintaining cell homeostasis by degrading and recycling biomolecules, regulating organelle quality control, and facilitating intracellular signaling. Lysosomal function is closely linked to the Golgi apparatus, endoplasmic reticulum, mitochondria, and nucleus, coordinating cellular metabolism and stress responses. When lysosomal function is impaired, damaged proteins and organelles accumulate, metabolic processes are disrupted, and cell membrane integrity is compromised, leading to various diseases. For example, in neurodegenerative diseases, lysosomal dysfunction leads to the accumulation of toxic aggregates, resulting in neuronal damage and cognitive decline. Understanding lysosomal regulation and its interactions with other organelles is critical for developing therapies to slow disease progression and promote cellular longevity.

 When conventional dyes are used to analyze lysosomal function, it is difficult to determine whether the lysosomal mass or their function (pH) has changed because the analysis is based only on the fluorescence intensity of a single dye.
 Dojindo's kits contain two types of dyes: pHLys Red/Green, which shows a lysosomal pH-dependent change in fluorescence intensity, and LysoPrime Green/Deep Red, which is lysosomal pH-resistant. By combining these two dyes, the lysosomal function can be analyzed in detail by simultaneously analyzing lysosomal mass and pH. 

Explore Dojindo's wide range of lysosomal staining and pH detection dyes. Choose the following kit or reagent that aligns with your experimental requirements.

Product Name (Item Code)	Supported Devices			Indicator and Detection Color	Dyes and Fluorescence Properties	Approximate Number of Use
Lysosomal Acidic pH Detection Kit-Green/Deep Red (L268)	✓	✓	✓	pH	pHLys Green Ex: 488 nm / Em: 490-550 nm	[for 1 set] 35 mm dish: 10 dishes μ-Slide 8 well: 10 plates 96-well Plate: 2 plates
				quantity	LysoPrime Deep Red Ex: 633 nm / Em: 640-700 nm
Lysosomal Acidic pH Detection Kit-Green/Red (L266)	✓	Need G/Y Laser G:532 nm Y:561 nm	✓	pH	pHLys Red Ex: 561 nm / Em: 560-650 nm
				quantity	LysoPrime Green Ex: 488 nm / Em: 500-600 nm
pHLys Red- Lysosomal Acidic pH Detection  (L265)	✓		✓	pH	pHLys Red Ex: 561 nm / Em: 560-650 nm	[for 1 tube] 35 mm dish: 10 dishes μ-Slide 8 well: 10 plates 96-well Plate: 2 plates
LysoPrime Deep Red - High Specificity and pH Resistance  (L264)	✓	✓	✓	quantity	LysoPrime Deep Red Ex: 633 nm / Em: 640-700 nm
LysoPrime Green- High Specificity and pH Resistance  (L261)	✓	✓	✓	quantity	LysoPrime Green Ex: 488 nm / Em: 500-600 nm	[for 10 μl] 35 mm dish: 10 dishes μ-Slide 8 well: 10 plates 96-well Plate: 2 plates

 

 

Endocytic vesicles were labeled by ECGreen and the lysosomal mass and pH were detected separately with LysoPrime Deep Red and pHLys Red. Co-staining with ECGreen and Lysosomal dyes showed the inhibition of endocytic vesicle-fusion induced by Bafilmycin A1.

CCCP and Antimycin are recognized inducers of mitochondrial ROS, linked to the loss of mitochondrial membrane potential. Recent studies have shown that CCCP induces not only mitochondrial ROS but also lysosomal dysfunction. To observe mitochondrial ROS, HeLa cells were labeled with MitoBright ROS Deep Red for Mitochondrial Superoxide Detection, and the lysosomal mass and pH were independently detected with LysoPrime Green and pHLys Red. Co-staining with MitoBright ROS and Lysosomal dyes revealed that CCCP, unlike Antimycin, triggers concurrent lysosomal neutralization and mitochondrial ROS induction.

Reference: Benjamin S Padman, et. al., Autophagy (2013)

Products in Use
   - LysoPrime Green
   - pHLys Red
   - Lysosomal Acidic pH Detection Kit
   - MitoBright ROS Deep Red - Mitochondrial Superoxide Detection

Related Products
   - Mitophagy Detection Kit and Mtphagy Dye

In neurodegenerative diseases, the relationship between lysosomal function and iron has attracted attention, and it has been reported^* that lysosomal neutralization prevents the breakdown of iron stores (Transferrin or Ferritin), resulting in a decrease in intracellular iron.
Lysosomal pH changes and intracellular iron changes in the same sample were detected using SH-SY5Y cells supplemented with lysosomal acidification inhibitor (Bafilomycin A1) or iron chelator (Deferipron (DFP)). (Lysosomal pH: Lysosomal Acidic pH Detection kit - Green/Deep Red, Intracellular iron: FerroOrange [Code:F374])
The results showed that the addition of Bafilomycin A1 decreased the fluorescence of FerroOrange, confirming the decrease in intracellular iron. The fluorescence of LysoPrime DeepRed remained almost unchanged, while the fluorescence of pHLys Green decreased due to lysosomal neutralization. These results suggest that there is a relationship between changes in intracellular iron and lysosome function.

*Mol Cell., 2020, 77(3), 645-655.

＜Condition＞
pHLys Green (Green) : Ex=488 nm, Em=486-574 nm
FerroOrange (Red) : Ex=561 nm, Em=550-650 nm
LysoPrime Deep Red (Violet) : Ex=633 nm, Em=599-700 nm