Multifunctional (3-in-1) cancer theranostics applications of hydroxyquinoline-appended polyfluorene nanoparticles

A biocompatible fluorescent hydroxyquinoline-affixed polyfluorene (PF-HQ) nanoparticle-based drug delivery system (DDS: PF-HQ–DOX) is utilized for cancer cell imaging, transport and delivery of the FDA-approved drug doxorubicin (DOX), and theranostics applications.


FT-IR, SEM and DLS measurement
For the identification of functional groups present in the nanomaterial (PF-HQ) and to know its

Biodistribution studies
To determine the bio-distribution of PF-HQ and doxorubicin in tumors and other vital organs in the treated C57BL6 mice, we collected all the required organs and tumors from the sacrificed

Immunofluorescence studies
Prior to immunostaining, the tumor tissue samples were fixed in ~ 4% PFA overnight and then washed with PBS for 2-3 times. 10 µm thicknesses of respective tissues were prepared by a Cryostat (Leica CM1950) and mounted on microscopic slides. The tumor tissue slides were dipped into isopropanol for 3-5 minutes followed by xylene for 3 minutes each and 10 mM citrate buffer (pH= 6.0) was used for antigen retrieval by heating sections in sub-boiling condition for 10 minutes.

Tunnel assay
To check the tissue apoptosis caused by PF-HQ-DOX, the fixed (4% formalin) tumor tissues were subjected to Tunel assay using an APO-DIRECT™ kit (BD Pharmingen, Cat. No. # 6536KK) according to the manufacturer's protocol. DAPI was used to tag the nucleus of the tumor cells. Images were acquired using a fluorescence microscope (Nikon Eclipse TE2000-E).
The green fluorescence emission (λ em = 525 nm) was collected after excitation at λ ex = 420-495 nm at 40X magnification. Similarly, the blue fluorescence emission (λ em = 485 nm) was collected after excitation at λ ex = 380 nm at 40X magnification. Figure S15. FT-IR spectra of Fluorenone (black), PF-HQ (red) and PF-DPA (blue). PF-HQ is presented in the present study. PF-DPA was explored in one of our previous studies. 1 No peak was found in the case of polymers at 1722 cm -1 .